Correlation between lncRNA GAS5 and LHPP expression and epithelial-mesenchymal transformation in non-small cell lung cancer tissues and clinical significance / 国际检验医学杂志

Objective To investigate the correlation between the expression of long non-coding ribonucleic acid growth arrest specific 5(lncRNA GAS5),phospholysine phosphohistidine inorganic pyrophosphate phos-phatase(LHPP)and epithelial-mesenchymal transition(EMT)in cancer tissues of patients with non-small cell lung cancer(NSCLC)and its clinical significance.Methods Cancer tissues and adjacent tissues of 90 patients with NSCLC who underwent surgical resection in the First Hospital Affiliated to Hebei North College from June 2018 to January 2020 were collected.The expressions of lncRNA GAS5,LHPP and EMT-associated pro-teins[E-calmodulin(E-Cad),N-calmodulin(N-Cad),and vimentin(VIM)]were detected by real-time fluores-cence quantitative polymerase chain reaction.The relationship between lncRNA GAS5 and LHPP mRNA and clinicopathological features in cancer tissues of NSCLC patients was analyzed,and the correlation between ln-cRNA GAS5 and LHPP mRNA and EMT-associated proteins expression in cancer tissues of NSCLC patients was analyzed by Pearson correlation.Kaplan-Meier method was used to plot the survival curves of NSCLC pa-tients with different lncRNA GAS5 and LHPP mRNA expressions,and multivariate Cox regression was used to analyze the prognostic factors of NSCLC patients.Results The expressions of lncRNA GAS5,LHPP mR-NA and E-Cad mRNA in cancer tissues of NSCLC patients were lower than those in adjacent tissues,while the expressions of N-Cad mRNA and VIM mRNA were higher than those in adjacent tissues,with statistical sig-nificance(P＜0.05).Pearson correlation analysis showed that lncRNA GAS5 in cancer tissues of NSCLC pa-tients was positively correlated with E-Cad mRNA expression(r=0.724,P＜0.001),and negatively correla-ted with N-Cad mRNA and VIM mRNA expression(r=-0.699,-0.689).P＜0.001);lncRNA GAS5 was positively correlated with LHPP mRNA expression(r=0.651,P＜0.001).The mRNA expressions of ln-cRNA GAS5 and LHPP in cancer tissues of NSCLC patients with different degrees of differentiation,tumor TNM stage and lymph node metastasis were significantly different(P＜0.05).Kaplan-Meier survival curve a-nalysis showed that the 3-year overall survival rate in the lncRNA GAS5 high expression group[68.18％(30/44)]was higher than that in the lncRNA GAS5 low expression group[36.96％(17/46)].The 3-year overall survival rate in the high LHPP mRNA expression group[67.39％(31/46)]was higher than that in the lowLHPP mRNA expression group[36.36％(16/44)],and the difference was statistically significant(x2=10.274,10.322,P＜0.05).Low differentiation,TNM stage Ⅲ and lymph node metastasis were independent risk factors for death in NSCLC patients,and lncRNA GAS5≥1.32 and LHPP mRNA≥1.12 were independ-ent protective factors(P＜0.05).Conclusion The low expression of lncRNA GAS5 and LHPP mRNA in cancer tissues of patients with NSCLC is related to EMT-associated proteins expression,differentiation de-gree,tumor TNM stage,lymph node metastasis and prognosis,and may become a new target for the diagnosis and treatment of NSCLC.

Efficacy of Hormone Combined with Cyclophosphamide in the Treatment of Connective Tissue Disease-associated Interstitial Lung Disease and Analysis of Its Influential Factors / 中国药房

OBJECTIVE：To investigate the efficacy of hormone combined with cyclophosphamide in the treatment of connective tissue disease-associated interstitial lung disease （CTD-ILD）and to analyze its influential factors. METHODS ：100 patients diagnosed as CTD-ILD in our hospital from Jan. 2018 to Jan. 2019 were randomly divided into observation group and control group ，with 50 cases in each group. Control group was treated with Compound cyclophosphamide tablets ，50 mg each time，3-4 times each day. Observation group was additionally treated with Prednisone acetate tablets ，10 mg each time ，3-4 times each day ，on the basis of control group. Treatment courses of 2 groups lasted for 6 months. The clinical efficacy ，the occurrence of ADR，lung function before and after treatment ，the levels of peripheral IL- 6，CRP and PCT and quality of life （SGRQ score ）were compared between 2 groups. According to the therapeutic efficacy ，all patients were divided into effective group and ineffective group. The related factors influencing the clinical efficacy of CTD-ILD were analyzed by univariate and multivariate Logistic regression analysis. RESULTS ：After treatment ，total response rate ，FVC，FEV1 and DLCO of observation group were significantly higher than those of control group ，while SGRQ score ，levels of IL- 6，CRP and PCT in peripheral blood were significantly lower than control group （P＜0.05）. There was no significant difference in the total incidence of ADR between 2 groups（P＞0.05）. Univariate analysis showed that there were no significant differences in gender ，age，past medical history and CTD type between effective group and ineffective group （P＞0.05）. However ，there were statistical significancant differences in the distribution of different levels of IL- 6，CRP and PCT in peripheral blood between 2 groups（P＜0.05）. Multivariate Logistic regression analysis showed that IL- 6 was an independent risk factor for therapeutic efficacy of combined therapy [OR （95％CI）＝ 4.537（3.668，10.352），P＝0.002]. CONCLUSIONS ：Hormone combined with cyclophosphamide can significantly improve the therapeutic efficacy of CTD-ILD patients ，improve their lung function and quality of life ，and reduce the expression level of inflammatory factors. The level of IL- 6 is an independent risk factor affecting the efficacy of the treatment ，and its changes should be paid close attention to during the treatment.

Diffusion-Weighted MRI for the Initial Viability Evaluation of Parasites in Hepatic Alveolar Echinococcosis: Comparison with Positron Emission Tomography

OBJECTIVE: More than 70% of hepatic alveolar echinococcosis (HAE) are inoperable. Thus, long-term, or even life-long, pharmacological treatment with benzimidazoles is necessary. For effective treatment, it is of great importance to employ imaging techniques to detect and monitor the non-resectable parasitic viability. Therefore, this study aimed to evaluate diffusion-weighted imaging (DWI) in assessing the viability of HAE in comparison to 18-fluoro-deoxyglucose (18F-FDG) positron emission tomography, combined with computed tomography (PET/CT). MATERIALS AND METHODS: Positron emission tomography, computed tomography and DWI (b-values: 0, 800 s/mm2) were retrospectively analysed in eight patients with clinically-verified HAE to, generate the apparent diffusion coefficient (ADC) map. The activity of HAE lesions in both techniques were determined independently by two radiologists according to the following standard: (+), marked focally or perilesionally increased FDG uptake/high signal intensity; (−), a hepatic defect without FDG uptake/no high signal intensity. Every lesion's maximum standardized uptake value (SUV(max)) on the PET/CT images and mean ADC values on the parametric ADC maps were measured respectively. Results of PET/CT and DWI were compared on a per-lesion-basis. Pearson's correlation coefficient was assessed for statistical analysis. RESULTS: A total of 14 HAE lesions were detected. Eight lesions (diameter 3–15 cm) showed perilesional hyper-signal intensity on DWI. This was visualised on PET/CT as increased FDG uptake. They mainly existed in the lesion's border with normal liver parenchyma. Five lesions (diameter < 2 cm) were detected as nodular hyperintensity on DWI and a ‘hot spot’ on PET/CT in the same distribution. One patient, who had received oral drug therapy for three years showed significantly decreased perilesional hyperintensity on the DWI and a hepatic defect without any FDG uptake on PET/CT. Pearson's correlation coefficient indicated a significant inverse correlation of the ADC and the SUV(max) (r = −0.67, p < 0.001). CONCLUSION: Diffusion-weighted imaging is capable of offering information on visually detecting the HAE lesions' viability and may be useful for routine application in the initial diagnosis of HAE.

Calcification characteristics and biological activity of hepatic alveolar echinococcosis / 中华放射学杂志

Objective To investigate the calcification characteristics and biological activity of hepatic alveolar echinococcosis(HAE). Methods Retrospective analysis of 60 patients in the First Affiliated Hospital of Xinjiang Medical University from June 2016 to April 2017 with hydatid positive or surgical pathology confirmed HAE.All patients underwent abdominal CT scan and double-phase enhanced PET-CT examination were confirmed with single lesion.The CT and PET-CT features were analysed,and the maximum standard uptake value (SUVmax) of the focal lesion on the PET-CT was measured. According to the calcification,HAE patients were divided into A,B,C type.Kruskal-Wallis H test was used to compare the difference of SUVmax between lesions of different HAE types. Results Sixty patients with hepatic alveolar echinococcosis were evenly divided into A,B and C by different calcifications.The median SUVmax of A,B and C were 3.41(2.17 to 3.75),7.45(6.77 to 9.01)and 6.67(6.28 to 9.01),respectively.The median SUVmax within three types was statistically significant (χ2=4.429, P<0.05). Conclusion The biological activity of different HAE calcifications is different.

Cancer-associated-fibroblasts regulate the chemoresistance of lung cancer cell line A549 via SDF-1 secretion / 中华肿瘤杂志

Objective@#To investigate whether cancer-associated- fibroblasts (CAF), the key component of tumor microenvironment, regulate the chemoresistant capacity of lung cancer cell line A549 through SDF-1 secretion.@*Methods@#Primary cell isolation techniques was used to isolate cancer-associated-fibroblasts from lung cancer patients. MTT assay was applied to determine the proliferation and chemoresistance of A549 cells. Quantative PCR was used to detect the mRNA changes of Bcl-xL. Western blotting was used to detect the protein expression of Bcl-xL. ELISA was applied to detect the SDF-1 secretion from normal fibroblasts (NF) and CAF.@*Results@#CAF promoted the proliferation of A549 cells, while NF had no significant effect on them. After 72 hrs incubation, the absorbance value of A549+ CAF medium group was 0.814±0.006, significantly different from the 0.753±0.006 of the A549+ NF medium group (P<0.05). The Q-PCR assay indicated that mRNA expressions of Bcl-xL in the A549 group, A549+ NF medium group and A549+ CAF medium group were 1.00±0.11, 1.10±0.09 and 3.50±0.30, respectively, showing a significant difference between the A549+ NF medium group and A549+ CAF medium group (P<0.05). The Western blot showed that protein expressions of Bcl-xL in the A549 group, A549+ NF medium group and A549+ CAF medium group were 1.00±0.08, 1.10±0.12 and 3.10±0.25, respectively, with a significant difference between the A549+ NF medium group and A549+ CAF medium group (P<0.05). The ELISA results showed that the SDF-1 concentrations in the A549+ NF medium group and A549+ CAF medium group were 3.23±0.02 and 9.53±0.10, respectively, significantly different from each other (P<0.05). The MTT assay indicated that the absorbance values of OD of A549 group, A549+ AMD3100 group, A549+ NF medium group, A549+ NF medium+ AMD3100 group, A549+ CAF medium and A549+ CA Fmedium+ AMD3100 group were 0.43±0.03, 0.25±0.02, 0.48±0.03, 0.31±0.03, 0.72±0.06 and 0.45±0.03, respectively. The data of A549+ NF medium group was significantly different from that of A549+ CAF medium group (P<0.05).@*Conclusions@#Cancer-associated-fibroblasts enhance the drug resistance of A549 cells through SDF-1 secretion, upregulating the expression level of Bcl-xL through interaction with CXCR4. Our study not only illustrates that tumor microenvironment is able to enhance drug resistance of tumor, but also provides experimental evidence for the cancer-associated-fibroblasts as a potential therapeutic target for the treatment of lung cancer.

Primary study on fusions of ovarian carcinoma cells to dendritic cell transfected with interleukin-12 gene in vitro / 中华妇产科杂志

Objective To investigate the immune response of the fusions of ovarian carcinoma cells to dendritic cell(DC) transfected with interleukin(IL)-12 gene in vitro. Methods Recombinant human granulocyte macrophage colony stimulating factor and recombinant human tumor necrosis factor ? were used to generate DC from cord blood in vitro. IL-12 fusion gene was cloned into a eukaryotic vector-pcDNA3.1(+). DC were transfected with IL-12-pcDNA3.1(+) using lipofectamine transfection method and electric transfection method respectively. Then polyethylene glycol mediated the fusion of transfected DC and ovarian carcinoma cells, and 3-(4,5-dimethylthiazol-2-yl) 2,5-diphenyl tetrazolium bromide (MTT) test was carried out to observe the immune response. Results DC were generated in vitro from cord blood and expressed high levels of costimulatory(50%) and MHC Ⅱ molecules(99%). RT-PCR showed that IL-12-pcDNA3.1(+) had been successfully transfected into DC. RT-PCR and immunofluorescence analysis showed that DC were fused with ovarian carcinoma cells successfully. Then the fusion cells of ovarian carcinoma cells to transfected DC, and the fusions of ovarian carcinoma cells to DC were cocultured with peripheral blood mononuclear cell respectively. MTT test showed that both fusions could induce cytolytic T cell activity and lysis of ovarian carcinoma cells,and the former effect was stronger. Conclusion The cytolytic T cell activity induced by the fusions of ovarian carcinoma cells to transfected DC is enhanced.