Establishment and identification of radioresistant nasopharyngeal carcinoma cell line / 中华放射肿瘤学杂志

Objective:To establish a radioresistant nasopharyngeal carcinoma cell line in vitro and provide experimental basis for further research of the molecular mechanism of radioresistance. Methods:A radioresistant cell line 5-8F-IR was established by dose gradient irradiation. Cell morphological changes were observed by optical microscope. The formation of colony was detected by colony formation assay. Cell viability was detected by CCK-8 assay. Cell proliferation was assessed by 5-ethynyl-2'-deoxyuridine (EdU) assay. DNA damage repair ability was measured by immunofluorescence staining and comet assay. Cell apoptosis and cycle were detected by flow cytometry. The expression levels of DNA damage related protein γH2AX and apoptosis related protein Caspase-3 were measured by Western blot.Results:The 5-8F-IR cells became longer than that of parental generation 5-8F cells after dose gradient irradiation. The colony formation ability ( P<0.01), cell viability ( P<0.001), cell proliferation ability ( P<0.05) and DNA damage repair ability ( P<0.05) of 5-8F-IR cells were significantly stronger than those of parental generation 5-8F cells. The apoptosis rate of 5-8F-IR cells after irradiation was significantly lower than that of parental generation 5-8F cells ( P<0.01). The 5-8F-IR cells showed higher percentage of cells in S phase without irradiation, and obvious G 2/M phase arrest after irradiation ( P<0.01). Western blot showed that the expression levels of γH2AX ( P<0.001) and Caspase-3 ( P<0.05) in 5-8F-IR cells after irradiation were significantly lower compared with those of parental generation 5-8F cells. Conclusion:5-8F-IR cells induced by dose gradient irradiation of human nasopharyngeal carcinoma cell line 5-8F cells exhibit radioresistance and exhibit different biological characteristics compared to their parental 5-8F cells, providing a research tool for exploring radioresistance mechanism of nasopharyngeal carcinoma.

Research progress on radio-resistance mechanism of nasopharyngeal carcinoma / 中华放射肿瘤学杂志

Nasopharyngeal carcinoma is one of the most common malignant head and neck tumors, and radiotherapy is the main treatment. However, radio-resistance is a key cause of local recurrence of nasopharyngeal carcinoma. Therefore, overcoming the radio-resistance of nasopharyngeal carcinoma and enhancing the radiosensitivity have become urgent problems in the treatment of nasopharyngeal carcinoma, which also play a key role in improving the overall survival rate of patients. In this article, recent studies on DNA, non-coding RNA (ncRNA), protein and cell behaviors related to radio-resistance of nasopharyngeal carcinoma were reviewed, aiming to provide valuable ideas for clinical treatment of nasopharyngeal carcinoma.

Effect of electro-acupuncture therapy on limb spasm and excitability of motor neurons in stroke rats / 浙江大学学报·医学版

To investigate the effect of electro-acupuncture therapy on limb spasm and excitability of motor neurons in stroke rats. Ischemic stroke model was induced with middle cerebral artery embolization in SD rats. Thirty-three modeled rats were randomly divided into model group, electro-acupuncture group, and baclofen group with 11 rats in each group, and another 10 rats were taken as sham operation group. The electro-acupuncture group and the baclofen group were treated with electro-acupuncture and baclofen tablets respectively. The model group and the sham operation group had no intervention. The neural function was evaluated with Bederson's scale and balance beam test; the muscle tension was measured with electrophysiography; the pathological changes of brain tissue was examined with HE staining; the content of glutamic acid (Glu) and γ-aminobutyric acid (GABA) in rat cerebral cortex was analyze with enzyme linked immunosorbent assay (ELISA) method, the expression of metabotropic glutamate receptor 1a () and γ-aminobutyric acid type B receptor subunit 1 () mRNA were detected with RT-qPCR. Compared with the model group, the neurological function scores of the electro-acupuncture group and the baclofen group showed a downward trend at d7 after operation (all >0.05), and the neurological function scores of the electro-acupuncture group and the baclofen group were significantly decreased at d12 after the operation (all 0.05). Compared with the model group, the electrophysiological results of the electro-acupuncture group and baclofen group were significantly increased after operation (all <0.05). The results of HE staining showed that there was no cell edema and degeneration in the sham operation group, no pyknosis of the nucleus, and no bleeding in the interstitium. Cell edema and degeneration and mesenchymal congestion appeared in the model group. Compared with the model group, the cytoplasmic edema and degeneration and the interstitial bleeding in the electroacupuncture group and the baclofen group were reduced. Compared with sham operation group, the Glu content and the relative expression of mRNA was increased in the model group, electro-acupuncture group and baclofen group, while the GABA content and the relative expression of mRNA decreased (all <0.05). Compared with model group, the Glu content and the relative expression of mRNA in the electro-acupuncture group and baclofen group decreased, and the GABA content and relative expression of mRNA increased (all <0.05). Electro-acupuncture may improve limb spasm after stroke through regulating the expression of Glu and GABA in the cerebral cortex and the excitability of motor neurons in rats.

LincRNA-ROR functions as a ceRNA to regulate Oct4, Sox2, and Nanog expression by sponging miR-145 and its effect on biologic characteristics of colonic cancer stem cells / 中华病理学杂志

Objective@#To investigate the impact of lincRNA-ROR, a ceRNA by binding miR-145 on the expression of the downstream genes Oct4, Sox2 and Nanog, and related biological characteristics of colon cancer stem cells, and to elucidate the clinical significance of this molecular regulatory network.@*Methods@#Fifty-two cases of colorectal cancer tissue and adjacent tissue were collected at Nanyang City Central Hospital and Nanyang Second Hospital, Henan Province, from 2014 to 2016. Real-time quantitative polymerase chain reaction (qPCR) was used to detect the expression of lincRNA-ROR and miR-145 in colorectal cancer tissue and isolated colon cancer cells. The correlation between the expression of lincRNA-ROR, miR-145 and the clinicopathologic features of colon cancer was performed. CD44-CD133- and CD44+ CD133+ cells were isolated from SW1116 by using flow cytometry. The expression of CD44, CD133, Oct4, Sox2, Nanog, lincRNA-ROR and miR-145 in cells were detected by qPCR. The relationship between lincRNA-ROR, miR-145, Oct4, Sox2 and Nanog was analyzed by bioinformatics, dual luciferase reporter assay, qPCR and Western blot. The effects of silencing lincRNA-ROR on the proliferation and chemosensitivity of colon cancer stem cells were detected by MTT, colony formation.@*Results@#LincRNA-ROR was frequently up-regulated and inversely correlated with miR-145 down-regulation in the colon cancer specimens(P<0.05). LincRNA-ROR was related to tumor size, lymph node involvement and distant metastasis(P<0.05), and miR-145 was found related to tumor size and tumor location(P<0.05). CD44+ CD133+ cells were successfully isolated from SW1116 by flow cytometry. The levels of CD44, CD133, Oct4, Sox2, Nanog, lincRNA-ROR in CD44+ CD133+ cells were significantly increased, while miR-145 was decreased compared with CD44-CD133-cells(P<0.05). The levels of CD44, CD133, lnc-ROR in CD44+ CD133+ cells were significantly reduced upon cell adherence, while miR-145 was significantly increased(P<0.05). Bioinformatics analysis revealed that lincRNA-ROR shared miRNA response elements with core transcription factors Oct4, Sox2 and Nanog. MiR-145 significantly inhibited the expression of lincRNA-ROR, Oct4, Sox2 and Nanog. Silencing lincRNA-ROR significantly inhibited colon cancer stem cells proliferation and increased the sensitivity to chemotherapy.@*Conclusions@#Linc-ROR functions as a key ceRNA to prevent core TFs, e. g., Oct4, Sox2, Nanog, from miR-145-mediated suppression in colon cancer stem cells and regulates cell proliferation and chemosensitivity.The data may provide insights into the pathophysiological interactions of the components of genetic networks in the development of colon cancer and may lead to new therapies in the future.

Research progress in drug-induced aplastic anemia / 中国综合临床

Aplastic anemia(AA)is a rare disease oringed from hematopoietic cell,characterized by failure of the bone marrow and pancytopenia in peripheral blood.The mechanism of AA is indistinct,it can be congenital,or acquired disposition.Acquired AA may be secondary to poisonous,radiation,virus infection and some medicine.All these factors can injure bone marrow directly,suppressing the proliferation and differentiation of bone marrow,on the other hand,the immune system play a pivotal role indirectly.This review focused on the pathologic classification,mechanism of drug-induced aplastic anemia.

microRNA profile changes in development of NKT cells / 中国免疫学杂志

Objective:To explore the microRNA profile changes in the development of NKT cells.Methods: Differently developmental stage of NKT cells in mouse thymus were sorted by flow cytometry.Total RNA were extracted,reversely transcribed and pre-amplified.TaqMan low density microRNA assay and single real-time PCR were applied to detect the expression changes of microRNAs in the developmental process of NKT cells.Results: There were total 92 microRNAs whose expression changed significantly during the development and maturation of NKT cells.Among them,increasly expressed microRNAs were 71,including 36 microRNAs whose expression continuously increased;decreasly expressed microRNAs were 21,including 12 microRNAs whose expression continuously decreased.In addition,single real-time PCR analysis showed that the expression of Let-7f,miR-150,miR-24,miR-29 increased,while the expression of miR-223 and miR-155 decreased during the development and maturation of NKT cells.Conclusion: NKT development and maturation is accompanied by expression changes of large amount of microRNAs,indicating that specific microRNA regulates NKT development and function.

Clinicopathological features, immunohistochemistry and differential diagnostic analysis of metaplastic breast carcinoma / 重庆医学

Objective To analyze the clinicopathological features,immunohistochemistry and differential diagnosis of metaplastic breast carcinoma.Methods A total of 32 cases of patients with metaplastic breast cancer in our hospital from March 2013 to March 2016 was randomly selected.The specimens were fixed with 4％ neutral formaldehyde solution,then dehydrated,embedded,sliced,stained with HE,and immunohistochemically stained with immunohistochemical staining machine,and then the results of macroscopic examination,microscopic examination and immunohistochemical examination were analyzed.Results Metaplastic breast cancer was lack of clear boundaries,and had no envelope.The tumor diameter was 1-5 cm,and its average was(3.1 ±1.2) cm.Section was tough and solid.Some had nodular shape,and their color was gray.Some had cysts appearance in the section.Among the 32 patients,20 patients underwent sentinel lymph node biopsy without cancer metastasis,and 12 patients underwent axillary lymph node dissection,including 8 cases with axillary lymph node metastasis;32 patients were presented as estrogen receptor (ER) (-),progesterone receptor(PR) (-),epidermal growth factor receptor-2 (HER-2) (-),broad spectrum CK (+),28 patients presented as epidermal growth factor receptor(EGFR)(+),20 patients presented as CK5/6 (+),and 8 patients presented as p53 (+),myoepithelial carcinoma p63 (+),respectively accounting for 100.0％,87.5 ％,62.5 ％ and 25.0％ in their total number.A total of 32 cases of patients with metaplastic breast carcinoma was divided into two types according to the metaplasia:pure epithelial type,including 4 cases of squamous cell carcinoma,2 cases of glandular squamous cell carcinoma and 13 cases of adenocarcinoma differentiated by spindle cell;epithelial/mesenchymal metaplastic carcinoma,its heterologous elements including 3 cases of sarcomatoid components,3 cases of rhabdomyosarcoma-like ingredients and 7 cases of ossification of cartilage and natural ingredients.Conclusion Metaplastic breast cancer,ER(-),PR(-),and HER-2 (-) should be identified with primary breast sarcoma,malignant follicular tumor,sarcomatosis and desmoplastic breast cancer.

Clinical significance of CUL4B and interleukin-6 levels in patients with multiple myeloma / 中国综合临床

Objective To investigate the clinical significance of CUL4B and interleukin?6( IL?6) levels in patients with multiple myeloma(MM).Methods The mRNA of CUL4B and IL?6 were detected by real?time relative quantitative PCR and enzyme?linked immunosorbent assay( ELISA) in 46 cases of MM and 40 cases of non hematologic cancer patients.Results CUL4B concentrations in patients with MM were 3.455(2.098,4. 768).IL?6 concentrations in patients with MM were significantly higher than those of the normal control group ((23. 985(23. 015,26. 878) ng/L vs. 6. 205(5. 405,10. 215) ng/L,Z=-8. 262,P=0. 000). The expression of CUL48 mRNA in Stage Ⅰ,Ⅱ,Ⅲ was 2. 424 ( 1. 881, 3. 583 ) , 3. 594 ( 2. 093, 5. 738 ) , 4. 300 ( 2. 928, 7. 272) respectively, and IL?6 was 23. 115 ( 22. 723, 23. 568 ) , 23. 630 ( 22. 860, 26. 625 ) , 26. 35 ( 24. 995, 30. 550) ng/L respectively,with the increase of clinical stage,the level of CUL4B and IL?6 showed increasing trend( P<0. 05) . CUL4B concentrations were significantly decreased in MM patients after treatment than before treatment(1. 665(1. 420,2. 298) vs. 3. 455(2. 098,4. 768),Z=-4. 955,P=0. 000). IL?6 concentrations were significantly decreased in MM patients after treatment than before treatment( 15. 160( 11. 705,17. 195) ng/L vs. 23. 985(23. 015,26. 878) ng/L,Z=-7. 981,P=0. 000). CUL4B level in patients was positively correlated with IL?6( r=0. 386,P=0. 008 ) . Conclusion It is of great significance to detect CUL4B and IL?6 in patients with MM on clinical stage,the condition of judgment and efficacy observation.

Clinical significance of Her2 pathological expression in breast and gastric cancer patients / 中国免疫学杂志

Objective:To investigate the clinical significance of Her2 pathological expression in the breast and gastric cancer patients for providing a reference for cancer prevention. Methods: Selected surgical resection specimens of 60 diagnosed in advanced gastric cancer and 60 diagnosed with advanced breast cancer from March 2009 to May 2014 in our hospital,all specimens were given the Her2 pathological expression of immunohistochemical analysis and investigation the clinical and pathological data. Results: The Her2 positive expression rates in the breast cancer and gastric cancer specimens were 40. 0% and 36. 7%,respectively that compared to no significant difference (P>0. 05). The Her2 expression was not related to the cancer patient’s age,histological type,and there were sig-nificantly correlated to the TNM stage and lymph node metastasis ( P<0. 05 ) . Spearman correlation analysis showed that Her2 expression in the breast cancer and gastric cancer were significant positive correlation to the Survivin, Bcl-2 expression ( P<0. 05 ) . Conclusion:Breast cancer and gastric cancer patients have shown high pathological expression of Her2,and are related to the TNM stage and lymph node metastasis, it may be through inhibited the apoptosis regulating proteins involved in the pathogenesis and metastasis of tumors.

A study on the effective factors of hospital acquired infections in chronic hepatitis B virus infection / 中国综合临床

Objective To investigate hospital acquired infections in patients with liver cirrhosis caused by relevant factors.Methods From July 2008 to June 2013,the clinical data of 470 cases of hospital acquired infections patients with chronic hepatitis B virus (HBV) infection were retrospectively analyzed by case-control study for the effective factors.Results Hospital acquired rate was 29.1％ (137/470).By logistic regression analysis,the effective factors were length of stay (OR =27.824,95％ CI 7.187-98.386),invasive operation (OR =17.201,95％ CI 4.245-71.303),a complication (OR =2.138,95％ CI 1.030-4.377),preventive use of antibiotics(OR =2.741,95％CI 1.816-4.010),drinking history(OR=34.248,95％CI 13.045-82.328),serum albumin(OR =17.258,95％ CI 6.242-53.162),quantitative PCR-HBVDNA (OR =4.859,95％ CI 3.214 -7.625),white blood cell (OR =4.271,95 ％ CI 1.520-12.157),c holinesterase (OR =2.761,95 ％ CI 1.523 -3.787) and anti virus medicine(OR=0.128,95％CI 0.041-0.375) (P＜0.05).Conclusion This study shows that length of stay,invasive operation history,complications,low white blood cell,low serum protein,low cholinesterase,the prophylactic use of antibiotics,hormone,high PCR-HBVDNA quantitative and drinking are the important risk of hospital acquired infections infection factors of patients of impact chronic HBV infection.Applications of antiviral drugs are effective in the prevention of chronic HBV infection protection factors of hospital acquired infections infection patients.

Clinical features and risk factors of silent cerebral infarction: an analysis of 138 patients / 中华神经医学杂志

Objective To explore the clinical features,risk factors and gender differences of silent cerebral infarction (SCI),and provide the clinical basis for the primary and secondary preventions.Methods Three hundred and fifty-six patients without previous history of stroke and neurological signs,admitted to our hospital from January 2010 to January 2013,were selected and divided to 2 groups:an observation group (n=138,having SCI) and a control group (n=218,not having SCI).Age,gender,levels of blood pressure,blood glucose,triglyceride,low density lipoprotein (LDL) and total cholesterol,smoking history,drinking history,family history of stroke,and coronary heart disease history were analyzed by single and multiple factor Logistic analysis.SCI patients were divided into male SCI group and female SCI group to compare the differences of high risk factors.Results SCI lesions mainly located in the basal ganglia and crown areas of radiation,which were mainly multiple lesions.Logistic regression analysis showed that independent risk factors were age,hypertension,coronary heart disease history,diabetes history,high triglyceride and high LDL levels,and smoking history.The incidences of diabetes and coronary heart disease in female SCI patients were significantly higher than those in male SCI patients (P＜0.05),and those of high triglyceride and smoking in SCI female patients were statistically lower than those in male SCI patients (P＜0.05).Conclusion Risk factors of SCI include age,hypertension,coronary heart disease history,diabetes,high triglyceride and high LDL levels,and smoking;and gender differences exist in these risk factors.

Treatment of human pancreatic cancer with adenovirus-mediated fusion gene system driven by KDR promoter in nude mice / 中华肝胆外科杂志

Objective To investigate the curative effect of the adenovirus-mediated fusion gene system driven by KDR promoter (AdKDR-CDglyTK) on a model of pancreatic cancer. Methods By using transplantation of the cultivated cells, human pancreatic cell line Capan-2 was injected subcutaneously on the back of nude mice to establish the animal model of the pancreatic cancer. Twenty nude mice were divided randomly and equally into four groups. The mice in group Ⅰ were injected with AdKDR-CDglyTK and 5-FC/GCV, those in group Ⅱ were injected with 5-FC/GCV, those in group Ⅲwere injected with AdKDR-CDglyTK and those in group Ⅳ received no any injection. AdKDR-CDglyTK was injected directly into the tumor and 5-FC/GCV was given by intraperitoneal injection. The observing parameters included common status, tumor bulk, tumor weight, inhibition rate of tumor growth, pathology, immunohistochemistry and treatment effect in each group. Electron microscopy was performed to observe the pathological changes of cells. The apoptotic cells in tumor were detected using the TUNEL assay. The expression of CDglyTK in tumors from each group was examined by RT-PCR. Results Tumor growth was dramatically inhibited in group Ⅰ. Tumor growth has no significant difference among groupⅡ , group Ⅲ and group Ⅳ. The apoptotic rate (34.20±4.60)% was significantly increased in group Ⅰ (F= 243. 22, P= 0. 00) and it had no significant difference among groupⅡ , group Ⅲ and group Ⅳ (P＞0.05). Conclusion AdKDR-CDglyTK with 5-FC/GCV can obviously inhibit the growth of human KDR-expressing pancreatic cell line Capan-2 and induce the cell apoptosis in vivo. The probable molecular mechanism lies in the facts that the system can cause a decline in the level of Bcl-2.

Adenovirus mediated fusion gene system driven by VEGF promoter kills pancreatic cancer cell / 中华普通外科杂志

Objective To evaluate the selectively killing effect of adenovirus(Ad)mediated double suicide gene driven by VEGF promoter on pancreatic cancer cell SW1990. Methods VEGFexpressing SW1990 were infected by Ad-VEGFP-CDTK and Ad-null.respectively.The infection rate was observed and the expression of CDTK was detected by RT-PCR and Western blotting.Followed by treatment with 5-FC and GCV killing effects were evaluated and bystander effects were analyzed by MTF.Pathological character of cells was observed by electron microscopy and distribution of cell cycle was detected by flow cytometry.The caspase-3 activity was detected by absorption spectrometry. Results The infection rate of the resultant recombinant Ad to SW1990 cells was not apparently different.RT-PCR and Western blotting demonstrated product of CDTK gene in SW1990 cell infected by Ad-VEGFP-CDTK.Prodrug could inhibit proliferation of SW1990 and the effect was dose-dependent.There was considerable bystander effect as observed by MTF.Apoptotic peak was also shown by flow cytometry.Morphologic features of apoptosis in SW1990 cells were displayed via electron microscopy.Cells at the G0-G1 phase was increasing and the rate at the G2-M and S phase was decreased by prodrug.The caspase-3 activity gradually rised with the increasing concentration of the prodrug. Conclusions The CDTK fusion gene system controlled by VEGF promoter has killing effect on the VEGF-expressing SW1990 cells and inducing the cell apoptosis.