Expression and Significance of Mitochondrial Autophagy-associated Protein PINK1 and Parkin in Parotid Pleomorphic Adenoma and Carcinoma ex Pleomorphic Adenoma / 肿瘤防治研究

Objective To investigate the expression of mitochondrial autophagy-associated protein PINK1 and Parkin in parotid pleomorphic adenoma (PA) and carcinoma ex pleomorphic adenoma(CA-EX-PA). Methods The expression of PINK1 and Parkin were detected by immunohistochemistry in 24 cases of normal parotid gland tissues, 32 cases of PA tissues and 42 cases of CA-EX-PA tissues. The correlations of PINK1 and Parkin expression with the clinicopathologic characteristics of CA-EX-PA patients were analyzed. Results The positive rates of PINK1 in normal parotid gland, PA and CA-EX-PA tissues were 100%, 91% and 67% respectively; and those of Parkin were 100%, 88% and 52% respectively. The expression rates of PINK1 and Parkin in PA and CA-EX-PA tissues were significantly lower than those in normal tissues (P < 0.05). The expression of PINK1 and Parkin protein were positively correlated in CA-EX-PA tissues (r=0.877, P < 0.01). In CA-EX-PA tissues, the expression of PINK1 and Parkin were associated with tumor invasion, lymph node metastasis and TNM stage (P < 0.05). Conclusion The expression of PINK1 and Parkin are decreased in PA and CA-EX-PA tissues. The decrease of mitochondrial autophagy activity might play important roles in the development, invasion and metastasis of parotid gland tumors.

Expression and clinical significance of phosphoenolpyruvate carboxykinas and plaminogen activator inhibitor-1 in pleomorphic adenoma and adjacent tumor tissues of parotid gland / 实用医学杂志

Objective To explore the expression and clinical significance of phosphoenolpyruvate carboxykinase (PCK) and plaminogen activator inhibitor-1 (PAI-1) in pleomorphic adenoma and adjacent tumor tissues of parotid gland. Methods Immunohistoehemistry was used to detect the expression of PCK and PAI-1 in10 cases of normal parotid glands, 38 cases of pleomorphic adenoma (0 cm) , adjacent tumor tissues (0.5, 1.0, 1.5, 2.0 cm from the pleomorphic adenoma). Results PCK expressed in all cases of normal parotid glands and adjacent tumor tissues, while PAI-1 expressed in 4 cases of normal parotid glands. Except 18 samples of 2.0 cm apart from pleomorphic adenoma, PAI-1 was expressed in the other adjacent tumor tissues. There was no obvious difference of PCK expression among normal, pleomorphic adenoma and adjacent tumor tissues of parotid glands (P> 0.05). The difference of PAI-1 expression was detected between pleomorphic adenoma (0 cm) and normal parotid glands and adjacent tumor tissues (2.0 cm from the tumor, P < 0.05). Conclusion During the development of pleomorphic adenoma, the expression level of PAI-1 is progressively increased while no change of PCK expression is found. It is rather remarkable that translocation of PAI-1 and PCK expression from cytoplasm and plasma membrane to nucleus occurs. The results mentioned above suggest that PCK and PAI-1 might play important roles in the progress of pleomorphic adenoma.

Correlations between Ape1/Ref-1, ICAM-1 and IL-17A Levels in Serum and Radiation Pneumonitis for Local Advanced Non-small Cell Lung Cancer Patients / 中国肺癌杂志

BACKGROUND@#The main manifestations of radiation pneumonitis are injury of alveolar epithelial and endothelial cells, abnormal expression of cytokines, abnormal proliferation of fibroblasts and synthesis of fibrous matrix. The occurrence of radiation pneumonitis is associated with multiplecytokine level abnormality. These cytokines can also be used as bio-markers to predict the occurrence of radiation pneumonitis. This study was to evaluate the correlation between the change of apurinic/apyrimidinic endonuclease 1/redox factor-1 (Ape1/Ref-1), intercellular adhesion molecules 1 (ICAM-1) and interleukin-17A (IL-17A) before and after radiotherapy and radiation pneumonitis for local advanced non-small cell lung cancer (NSCLC) patients with concurrent chemoradiotherapy.@*METHODS@#NSCLC patients (68 cases) were treated with concurrent radiotherapy and chemotherapy, every patient's normal tissue were controlled with a same radation dose. 68 local advanced NSCLC patients with concurrent chemoradiotherapy were detected the levels of Ape1/Ref-1, ICAM-1 and IL-17A in serum by ELISA before radiotherapy and in the 14th week after radiotherapy. Acute and advanced radiation pulmonary injury was graded according to Radiation Therapy Oncology Group/European Organization For Research and Treatment (RTOG/EORTC) diagnostic and grading criteria. Grade 2 or more radiation pneumonitis was taken as the main end point.@*RESULTS@#Eighteen cases out of 68 developed radiation pneumonitis, 50 of 68 cases have no radiation pneumonia development. There was no significant change of Ape1/Ref-1 levels before and after radiotherapy in radiation pneumonitis group (P>0.05). There was no significant change of Ape1/Ref-1 concentration in serum after radiotherapy between radiation pneumonitis group and non-radiation pneumonitis group (P>0.05). Compared with before radiotherapy, upregulation degree of ICAM-1 levels in radiation pneumonitis group was significantly higher than that in non- radiation pneumonitis group (P<0.05). There was no significant change of IL-17A concentration before and after radiotherapy in radiation pneumonitis group, but after radiotherapy IL-17A concentration in serum were remarkably higher than that in non-radiation pneumonitis group (P<0.05). Correlation analysis found that the change of ICAM-1 before and after radiotherapy has no obvious correlation with the incidence of radiation pneumonitis, and IL-17A change has obvious correlation with the incidence of radiation pneumonitis.@*CONCLUSIONS@#On the basis of strictly controlling radiation dose on normal tissue, IL-17A in serum could be the predictive factors of radiation pneumonitis for local advanced NSCLC patients with concurrent chemoradiotherapy.

Value of introvoxel incoherent motion imaging on evaluating concurrent chemoradiotherapy responseinpatientswithadvanceduterinecervixcancer / 中华放射学杂志

Objective To investigate the value of introvoxel incoherent motion(IVIM)using 3.0 T MRI to evaluate response to concurrent chemoradiotherapy(CCRT)in patients with advanced uterine cervix cancer. Methods From July 2015 to December 2016,63 patients with advanced(≥ⅡB)cervical cancer diagnosed by clinical and imaging study, who had completed CCRT plan in Henan Cancer Hospital, were prospectively enrolled.Pelvic MRI protocol including T1WI,T2WI,IVIM and dynamic contrasted enhanced scans were performed in each patient before CCRT and 3 weeks after starting therapy(total dose of 30 Gy), and at the end of therapy (total dose of 90 Gy, 8 weeks after therapy). The mean values of ADC, true molecular diffusion coefficient(D),pseudodiffusion coefficient(D*)and perfusion fraction(f)in each tumor at pre-therapy, in the middle of therapy and post-therapy were measured and recorded as ADC-pre, D-pre, D*-pre,f-pre;ADC-mid,D-mid,D*-mid,f-mid and ADC-post,D-post,D*-post,f-post,respectively;the change rates of these parameters during and after therapy (recorded as ΔADC-mid, ΔD-mid, ΔD*-mid, Δf-mid;ΔADC-post, ΔD-post, ΔD*-post, Δf-post) were also calculated. Patients were classified into response group and non-response group,according to response evaluation criteria in solid tumors after CCRT.MRI imaging study was performed in each patient within 1 month after CCRT to follow up,and tumor regression rate was calculated.The Mann-Whitney U test was used to compare differences of parameters and their change rates between response group and non-response group. Spearman correlation analysis was performed to assess relationships between parameters, parameter change rates and tumor regression rate. Logistic regression model was applied to find potential ADC values for predicting therapeutic response. ROC was used to analyze efficacy of ADC values for evaluating therapeutic response in advanced uterine cervix cancer after CCRT. Results The mean value of tumor maximum diameter before and after therapy was (47.5 ± 12.9) and(12.8 ± 10.0)mm,tumor regression rate was(66.7 ± 33.6)%.Forty-eight patients were in the response group and 15 in the non-response group.The mean value of ADC-pre,D-pre,D*-pre and f-pre was 0.74(0.43, 1.14)×10-3,0.58(0.33,0.91)×10-3,12.12(2.30,21.4)×10-3mm2/s,9.65%(4.45%,13.89%),respectively.Tumor regression rate had positive correlation with ADC-pre and D-pre (r=0.773,0.840;P<0.05). Responders had increased ADC-pre, D-pre values than non-responders, which had statistically significant difference (P<0.05). Responders had increased ADC-mid, D-mid and f-mid values than non-responders, which had statistically significant difference (P<0.05), tumor regression rate had positive correlation with ADC-mid, D-mid and f-mid (r=0.808,0.834,0.563;P<0.05). Responders had increased ADC-post, D-post and f-post values than non-responders,which had statistically significant difference(P<0.05),tumor regression rate had positive correlation with ADC-post and D-post (r=0.799, 0.829;P<0.05).Tumor regression rate had positive correlation with ΔADC-mid,ΔD-mid,Δf-mid(r=0.526,0.573,0.454;P<0.05)and with ΔADC-post,ΔD-post, Δf-post (r=0.541, 0.555, 0.388;P<0.05). Responders had increased ΔADC-mid, ΔD-mid, Δf-mid and ΔADC-post, ΔD-post, Δf-post, which had statistically significant difference (P<0.05). Logistic regression analysis revealed only ADC-pre and D-post could be independent factors to predict therapeutic response in advanced uterine cervix cancer after CCRT,values of B,Wald,odds ratio and P was 22.488,8.431,1.429, 0.004 and 16.542,8.517,1.779,0.004.ROC analysis showed the area under the curve(AUC)of ADC-pre, D-pre,ΔADC-mid,ΔD-mid,Δf-mid,ΔADC-post,ΔD-post and Δf-post for predicting therapeutic response in advanced uterine cervix cancer after CCRT were 0.890,0.926,0.942,0.851,0.803,0.929,0.951 and 0.906, respectively. Conclusion The IVIM parameters before and during CCRT process and their changes are valuable for predicting and evaluating therapeutic response in advanced uterine cervix cancer after CCRT, with high clinical practice value.

Clinical features of infantile organic acidemia accompanied with acute metabolic crisis / 中国小儿急救医学

Objective To investigate the clinical and laboratory characters of infantile organic acidemia(OA) accompanied with acute metabolic crisis.Methods We analyzed retrospectively datum of infants with OA diagnosed in our unit from April 2006 to October 2014.Results Fity-three cases(37 male and 16 female,aged under 1 year old) were enrolled in this study,in which,28 cases were methylmalonic acidemia,11 cases were propionic acidemia,3 cases were biotinidase deficiency,3 cases were glutaric acidemia type Ⅱ,2 cases were glutaric acidemia type Ⅰ,2 cases were isovaleric acidemia,1 case was variety of coenzyme A carboxylase deficiency,1 case was glycerol kinase deficiency,1 case was 3-methylcrotonyl-CoA carboxylase deficiency and 1 case was holocarboxylase synthetase defect.Tweny-five of the 53 cases(47.2％)developed metabolic crisis within 7 days after onset,main manifestations included feeding difficulties,frequent seizures dyspnea,et al.The most common abnormal laboratory findings manifested severe hypoglycemia,intractable metabolic acidosis,hyperammonemia,et al.Twenty cases had family history.All patients were given symptomatic relief and supportive treatment,including colleting hypoglycemia,reducing hyperammonemia,keeping water,electrolyte and acid base balance,maintaining function of vital organs,suppling metabolic cofactor and special fomula,et al.After treatment,32 cases (60.4％) improved markedly while death occurred in 15 cases(28.3％).Conclusion OA confirmed under 1 year old is especially vulnerable to acute metabolic crisis,which characterized by sudden onset,rapid progress and is difficult to treat.Feeding difficulties,frequent seizures and dyspnea were the very common presentations.Early diagnosis and timely treatment are critical for improving the prognosis.Clinicians should be aware of it,and an early metabolic disorders screening should be intervened in patients with hypoglycemia of unknown etiology or refractory metabolic acidosis.

~(99)Tc~m-MIBI SPECT in prediction of response to chemotherapy in patients with small cell lung cancer / 中国综合临床

Objective To assess the prediction value of technetium-99m methoxyisobutylisonitrile(~(99)Tc~m-MIBI) for the effect of chemotherapy in small cell lung cancer (SCLC) patients. Methods Fifty-three patients with SCLC were divided into two groups according to the chest computed tomography (CT) examination, 39 patients in group A with complete or partial remission, and 14 patients in groups B with stable or progressive status. ~(99)Tc~m-MIBI was performed before chemotherapy. Following i. v. administration of 740 MBq ~(99)Tc~m-MIBI, SPECT imagings at 10 -30 minutes (early) and 2 -3 hours (delayed) were performed to obtain the uptake ratio of early phase tumor/normal lung tissue (ER) and the uptake ratio of delayed phase tumor/normal lung tissue (DR). The retention index (RI) was calculated as (DR-ER)/ ER × 100%. The differences of ER,DR and RI between the two groups were tested through t-test and rank sum test. Results ~(99)Tc~m-MIBI uptake was significantly higher in group A than group B: 2.33(SD:0.21) vs 2.02(SD:0.31) for the early ratio (ER) (t = -3.401, P<0.05) and 2.44(SD:0.19) vs 1.86 (SD :0.30) for the delayed ratio (DR) (t = - 6.724,P < 0.05). The median of RI in group A was signifi-cantly higher than that in group B (5.31% vs -9.26%,P <0.05). Conclusions ER,DR and RI of ~(99)Tc~m-MIBI SPECT may be helpful in predicting the response to chemotherapy in patients with SCLC.

Establishment and identification of stable cell line secreting humen IgE Cε-4 protein / 中华微生物学和免疫学杂志

Objective To establish a stable cell line secreting human IgE Cε2-4 protein, and in-vestigate the binding capacity of receptor FcεR Ⅰ Methods The E24 gene was derived from SKO-O07 cell line, and was then cloned into pcDNA3.1 (+) (signal peptides were synthesized and fused at the 5'-end of E24 gene) or pCMV-L vector. After transient transfection into 293T cell, the secreted F24 protein was ana-lyzed by sandwich ELISA. The best vector was chosen to be transfected into CHO cells with LipofectAMI-NETM 2000 reagent. After being selected by G418 and subcloned three times by limited-dilution method, two stable cell lines were established. E24 gene was amplified by RT-PCR, and the E24 protein in the superna-tant was identified by ELISA. Besides, the binding capacity of FceR ⅠⅡ was analyzed by flow cytometry method. Results Three mammalian expression vector SP-E24-F3. 1, SP lI-E24-P3.1 and E24-PL were constructed and transient transfected to 293T cells. The output of E24 protein in the supernatant were 19.1, 19.4 and 8.7 μg/ml, respectively. Then the vector SP IX-E24-P3.1 was transfected into CHO cells. Final-ly, two single clones secreting E24 protein were stably obtained. The output of E24 were all at least 25 μg/ml. RT-PCR could detect the E24 gene from one of the two clones. Furthermore, flow cytometry results showed that E24 could bind the receptor in a dose-dependent manner. Conclusion Two stable cell line se- creting E24 protein were obtained, while E24 could specifically bind FcεR Ⅰ.

A novel designed single domain antibody on 3-D structure of ricin A chain remarkably blocked ricin-induced cytotoxicity.

Ricin A chain (RA), an N-glycosidase, is able to fatally disrupt protein synthesis by attacking the Achilles heel of the ribosome RNA (rRNA). As specific immunotoxins, emergence of inhibitors for RA may obtain access to antagonistics against ricin intoxication and contribute to ameliorate the concomitant side effects. Many experimental results showed that the engineered VHs, which possessed solubility, stability, small size and consequently easier to express, purify and manipulate in vitro, were self and long-lived molecules compared to synthetic peptides. In this study, based on the crystal structure of RA, a novel recombinant human single-domain antibody expressing a polypeptide against RA in the CDR3 loop (named rVH(PT)) was obtained using computer-guided molecular design method. Theoretically, rVH(PT) could penetrate deeply into the active cleft of RA and act as a potent antagonist analogue to block the RA-rRNA interaction. Followed results showed that the recombinant VH(PT) was easily expressed of high-yield production and in a partially soluble fusion form in Escherichia coli. In vitro cytotoxicity experiments demonstrated that it possessed remarkable ability to block ricin-induced cytotoxicity. This study highlights the potential of human VH to display biostructure and biofunction of peptides designed on RA functional domain and could be useful in developing new antidotes with potential therapeutic uses to neutralize unintended exposure to ricin.