Identification of the glycosylation sites of Opsin3 and its glycosylation modification function / 生物工程学报

Opsin3 (OPN3) is a photoreceptor membrane protein with a typical seven-alpha helical transmembrane structure that belongs to the G-protein-coupled receptor (GPCR) superfamily and is widely expressed in brain. In recent years, it has been reported that OPN3 is also highly expressed in adipose tissue, and the protein is associated with the production of skin melanin. We found that the N82 site is the glycosylation site of OPN3. SNAP-tagTM has diverse functions and can be applied to a variety of different studies. By constructing a SNAP-tagged OPN3 recombinant protein, the distribution position of SNAP-OPN3 in cells can be clearly observed by fluorescence confocal microscopy using SNAP-Surface® 549 and SNAP-Cell® OregonGreen®, which provides a new method for studying the function of OPN3. It also shows that SNAP-tag does not affect the function of OPN3. Using the SNAP tag we found that OPN3 cannot be taken up to the cell membrane after glycosylation site mutation.

Prevalence analysis of noninvasive follicular thyroid neoplasm with papillary-like nuclear features in a Third Class A Level Hospital in Guiyang / 中华内分泌代谢杂志

Objective To analyze of the prevalence of encapsulated follicular variant of papillary thyroid carcinoma (EFVPTC) and noninvasive follicular thyroid neoplasm with papillary-like nuclear features (NIFTP) by 2017 World Health Organization ( WHO) classification of tumors of endocrine organs in Guiyang. Methods A retrospectively analysis of patients who had a thyroid surgery and confirmed thyroid cancer by pathological diagnosis in the Affiliated Hospital of Guizhou Medical University from 2009 to 2016. PTC and FVPTC by 2014 WHO classification of tumors of endocrine organs , and according to the 2017 WHO classification of tumors of endocrine organs ,the thyroid papillary carcinomas were reviewed and some had been confirmed as EFVPTC and NIFTP, with analysis of the prevalence and prognosis of NIFTP. Results Of the 1207 cases of thyroid carcinoma, 1150 cases were papillary carcinoma, the constituent ratio of thyroid carcinoma was 95.28％, the FVPTC was 72 cases, and the proportion of thyroid carcinoma was 5. 97％; the proportion of FVPTC in thyroid carcinoma decreased from 2009 to 2016 ( P <0.05). After pathological sections being reviewed, 10 cases had been confirmed as non-encapsulated infiltrative FVPTC, the ratio of thyroid cancer to thyroid cancer was 0.83％; EFVPTC was found in 62 cases, accounting for 5.14％ of thyroid carcinoma, these included 2 cases of NIFTP confirmed by the 2017 WHO classification of tumors of endocrine organs and 60 cases encapsulated invasive FVPTC, the percentage of thyroid carcinoma was 0.17％ and 4.97％. 62 cases of EFVPTC with the exception of 11 cases without further visit, while the remaining 51 cases of EFVPTC were followed up. Death, local or distant metastases were defined as adverse events. An adverse event was seen in 11 of 49 of the cases of invasive EFVPTC, including 2 died of disease; The NIFTP were alive with no evidence of disease. Conclusion The diagnosis of NIFTP according to new WHO classification of endocrine organ tumors in 2017 has little expected impact in Guiyang.

Expression of miR-125a-5p in diffuse large B-cell lymphoma and its significance / 实用医学杂志

Objective To investigate the expression of miR-125a-5p in diffuse large B-cell lymphoma and the relationship among the levels of NF-κB/p65,Bcl2,Caspase3 and Ki-67 protein in clinicopathological features, and the potential prognostic value. Methods A total of 84 cases of DLBCL were enrolled in this study. At the same time,10 cases of reactive hyperplasia tissues were used as the control group. The expression of miR-125a-5p was examined by SYBR Green real-time reverse transcriptase polymerase chain reaction(real-time RT-PCR). The expression of NF-κB/p65,Bcl2,Caspase3,Ki-67 was detected by the immunohistochemical staining assay. The clinical pathological data was collected and some patients were followed up. The clinical data was used for analysis of the prognosis of patients. Results MiR-125a-5p was shown up-regulated in the DLBCL samples ,which was (4.99 ± 8.22)times of that in the reactive hyperplasia samples(P=0.002). The up-regulation of miR-125a-5p in DLBCL was correlated with non-GCB subtype(P=0.027). The up-regulation of miR-125a-5p in DLBCL was sig-nificantly correlated with the expression of NF-κB/p65,Bcl2 and Caspase3(P=0.031,0.030,0.022). There was a significant correlation between non-GCB subtype and NF-κB/p65 level in DLBCL(P=0.002). Kaplan-Mei-er survival analysis showed that the high expression of miR-125a-5p was associated with the prognosis of DLBCL pa-tients(P=0.038). Multivariate Cox regression analysis showed that the high expression of miR-125a-5p may be an independent risk factor for the prognosis of DLBCL(P=0.013). Conclusions MiR-125a-5p may play an im-portant role in the diagnosis of DLBCL. The up-regulation of miR-125a-5p in DLBCL may affect the activation of NFκB/p65,especially in the non-GCB subtype. The up-regulation of miR-125a-5p in DLBCL may influence the de-velopment of tumor via apoptosis pathway. The expression of miR-125a-5p is associated with the poor prognosis of patients,which can be regarded as an independent risk factor for the prognosis of DLBCL.

Expression of miR-125a-5p in diffuse large B-cell lymphoma and its significance / 实用医学杂志

Objective To investigate the expression of miR-125a-5p in diffuse large B-cell lymphoma and the relationship among the levels of NF-κB/p65,Bcl2,Caspase3 and Ki-67 protein in clinicopathological features, and the potential prognostic value. Methods A total of 84 cases of DLBCL were enrolled in this study. At the same time,10 cases of reactive hyperplasia tissues were used as the control group. The expression of miR-125a-5p was examined by SYBR Green real-time reverse transcriptase polymerase chain reaction(real-time RT-PCR). The expression of NF-κB/p65,Bcl2,Caspase3,Ki-67 was detected by the immunohistochemical staining assay. The clinical pathological data was collected and some patients were followed up. The clinical data was used for analysis of the prognosis of patients. Results MiR-125a-5p was shown up-regulated in the DLBCL samples ,which was (4.99 ± 8.22)times of that in the reactive hyperplasia samples(P=0.002). The up-regulation of miR-125a-5p in DLBCL was correlated with non-GCB subtype(P=0.027). The up-regulation of miR-125a-5p in DLBCL was sig-nificantly correlated with the expression of NF-κB/p65,Bcl2 and Caspase3(P=0.031,0.030,0.022). There was a significant correlation between non-GCB subtype and NF-κB/p65 level in DLBCL(P=0.002). Kaplan-Mei-er survival analysis showed that the high expression of miR-125a-5p was associated with the prognosis of DLBCL pa-tients(P=0.038). Multivariate Cox regression analysis showed that the high expression of miR-125a-5p may be an independent risk factor for the prognosis of DLBCL(P=0.013). Conclusions MiR-125a-5p may play an im-portant role in the diagnosis of DLBCL. The up-regulation of miR-125a-5p in DLBCL may affect the activation of NFκB/p65,especially in the non-GCB subtype. The up-regulation of miR-125a-5p in DLBCL may influence the de-velopment of tumor via apoptosis pathway. The expression of miR-125a-5p is associated with the poor prognosis of patients,which can be regarded as an independent risk factor for the prognosis of DLBCL.

Vincristine reduces A20 siRNA-induced proliferation in diffuse large B cell / 基础医学与临床

Objective To investigate the impact of MDR1-targeting small interfering RNA (siRNA) on diffuse large B-cell lymphoma cell OCI-LY1 proliferation.Methods A20 gene was silenced using RNA interference.An optimal concentration and treatment duration of vincristine were selected using MTT.Before and after siRNA transfection, proliferation of OCI-LY1 cells was assayed using MTT assay, and cellular apoptosis was detected using FCM before or after the treatment of the cells with VCR.Detection of A20, NF-κB (p65) and Pgp proteins were conducted using Western blot whereas mRNA of the A20 and MDR1 genes were examined using real time PCR.Results1)Proliferation of OCI-LY1 cells was enhanced (P<0.001) after the transfection with siRNA-2,(P<0.05).In addition, cell proliferation curve was declined after VCR stimulation, but the decrease was slower in siRNA-transfected cells than the untransfected counterparts.2)Apoptostic rate was lower in siRNA-transfected cells than theuntransfected counterparts, and the rate was higher in the cells after treatment with the drug for 24 h (P<0.05).Increased apoptosis was more obvious in control OCI-LY1 cells than in siRNA-transfected cells after treatment with VCR(P<0.05).3)The expression of MDR1 mRNA and Pgp (P<0.001) was significantly increased after transfection, but the expression of MDR1 mRNA and Pgp were significantly decreased (P<0.05).The expression in VCR group was significantly lower than that in siRNA-transfected cells+VCR group (P<0.01).ConclusionsA20 siRNA could effectively enhance NF-kappa B expression in OCI-LY1 cells.NF-kappa B may up regulate the expression of its downstream genes such as MDR1 and cause apoptosis, in turn enhancing the inhibition of cell proliferation.VCR can reduce MDR1 mRNA and Pgp expression in OCI-LY1 cells and the effect of VCR could be attenuated by A20 siRNA.

Impacts of A20 gene deletion on clinicopathological features and prognosis of diffuse large B cell lymphoma and relative molecular mechanism / 重庆医学

Objective To detect the A20 gene deletion,investigate the impacts of A20 gene deletion on clinicopathological features and prognosis of DLBCL,and relationship between activation of NF-κB pathway and relative molecular pathogenesis.Methods A20 gene deletion was detected by fluorescence in situ hybridization (FISH).The expression of A20,Survivin,P65 and Ki-67 were detected by immunohistochemistry stain.Apoptosis was assayed by TUNEL.Follow-up and statistical analysis were done.Results The deletion rate of A20 gene was 21.7％.The deletion rate of A20 gene was obviously higher in ABC-like DLBCL than that in GCB-like DLBCL (30.6％ vs.8.3％,P＜0.05).It was observed that there was a negative correlation between A20 protein expression and A20 gene deletion (r=-0.259,P =0.023).The expression of P65 and Survivin protein was positively correlated with the A20 gene deletion (r=0.280,P =0.015;r =0.313,P =0.007).Apoptosis rate was significantly reduced in DLBCL patients with A20 gene deletion.The apoptosis rate was higher in cases with positive expression of A20 protein,while that was lower in cases with positive expression of p65 and Survivin protein than those with negative expression of corresponding protein.There was no statistically significant difference in apoptosis rate between ABC-like and GCB-like DLBCL patients (P＞0.05).COX regression analysis indicated that age,A20 gene deletion,types of DLBCL and Ki67 expression were independent factors associated with survival status.Log-rank test showed that there was a statistical difference in survival status between the cases with and without A20 gene deletion (P=0.015).Conclusion A20 gene deletion may associate with the attenuation of A20 protein expression.The latter weakens negative feedback regulation of A20 protein for NF-κB pathway.An up-regulated expression of Survivin and abnormal proliferation and apoptosis may be result from the abnormal activation of NF-κB.A20 gene deletion brings certain influence on clinical course and prognosis of DLBCL.

Effects of IL-6 and AG490 on Burkitt lymphoma cell growth / 中国病理生理杂志

AIM:To observe the influences of IL-6 and AG490 on the growth of Raji cell line ( Burkitt lym-phoma cell, BL).METHODS:Raji cells were cultured.IL-6, an activator of signal transducer and activator of transcrip-tion 3 (STAT3), and AG490, a specific inhibitor of STAT3 were added into the medium respectively .The expression of STAT3 and survivin at mRNA and protein levels was detected by real-time PCR and Western blot .The cell viability was measured by MTT assay .Apoptosis and cell cycle were examined by flow cytometry .RESULTS:IL-6 or AG490 affected the growth of Raji cells significantly in a dose-dependent manner (P<0.05).The mRNA expression of STAT3 and sur-vivin in Raji cells was higher in IL-6 group, and lower in the AG490 group than that in the corresponding control group . The statistical differences were found in the mRNA expression of STAT 3 and survivin among different IL-6 or AG490 groups (P<0.05).The concentration dependent relationship was also presented in IL-6 and AG490 groups by the regression a-nalysis.The results of Western blot showed that the protein levels of phosphorylated STAT 3 (p-STAT3), STAT3 and sur-vivin were increased in IL-6 group, and decreased in AG490 group.The apoptotic rate of Raji cells was gradually reduced with the increasing concentration of IL-6.The opposite results were detected in the Raji cells treated with AG 490.There was significant difference in constitute of the cell cycle between the groups treated with IL -6 or AG490 and corresponding control group.The cells at G1-phase and G1/S were significantly increased , while those at S-phase had no obvious change under treating with AG490.The cells at S-phase decreased obviously in the Raji cells treated with IL-6.CONCLUSION:IL-6 and AG490 distinctly affect the growth of Raji cells .The mechanism may be associated with the activation of STAT 3 and survivin.

The expression and clinical significance of p53,C-myc and CerbB-2 in thyroid papillary carcinoma / 重庆医学

Objective To investigate the expression level and clinical significance of p53 ,C-myc and CerbB-2 in papillary thy-roid carcinoma patients .Methods The expression level of p53 ,C-myc and CerbB-2 were detected in 45 cases of papillary thyroid carcinoma tissue specimens and 45 cases of nodular goiter tissue specimens by using immunohistochemical staining .Their correla-tions with clinic pathological features in papillary thyroid carcinoma were analyzed further .Results The positive expression rates of p53 ,C-myc and CerbB-2 in the papillary thyroid carcinoma tissue specimens were significantly higher than that of the nodular goiter tissue specimens(all P< 0 .01) .The expression of C-myc and CerbB-2 significantly correlated with lymph node metastasis(P< 0 . 05) .The expression of CerbB-2 significantly correlated with tumor size ,local infiltration ,TNM stage ,differentiated degree and lymph node metastasis (all P< 0 .05) .p53 positive expression ,C-myc positive expression and CerbB-2 positive expression were the independent factors of cervical lymph node metastasis .Conclusion p53 ,C-myc and CerbB-2 were highly expressed in papillary thy-roid carcinoma and correlated with cervical lymph nody metastasis .The detection of p53 ,C-myc and CerbB-2 might be helpful for diagnosis of papillary thyroid carcinomas .

Relationship of A20 mutation with clinicopathologic features and prognosis of diffuse large B cell lymphoma / 中国肿瘤临床

Objective:To detect A20 mutation and to investigate its relationship with clinicopathologic features, prognosis, and re-sistance to therapy of diffuse large B cell lymphoma (DLBCL). Methods:A total of 104 cases with DLBCL and their clinical data were collected;follow-up was performed on a few of DLBCL patients. The expression of P-gP and Ki-67 protein was detected with immuno-histochemical staining. The A20 gene mutation in exons 3, 6, and 7 were examined by polymerase chain reaction and DNA sequencing. Finally, the correlation of genetic abnormality of A20 with clinicopathologic features was analyzed. Results:Missense mutation in ex-on 3 of A20 gene was identified in 18 of 104 patients (17.3%). The A20 gene mutation at site 73 of exon 3 was greater in the cases with activated B cell-like-DLBCL than with germinal center B-cell-like-DLBCL (18.5%vs. 2.5%, P=0.010). In contrast to the advanced clin-ical stage and high International Prognostic Index (IPI) cases, the rate of A20 mutation was superior to the opposite (P<0.05). The ex-pression for P-gP was higher in the cases with mutation than that of those with wild-type A20 gene (16/18 vs. 52/86, P=0.021). The dif-ference in A20 mutation between the groups of low and high positive expression for Ki-67 (1/17 vs. 26/60, P=0.030) was significant. DLBCL with A20 mutation had an increasing recurrence tendency (P=0.06) and a worse survival (P=0.016) compared with those with wild-type A20 gene. Conclusion:The A20 mutation has a certain influence on the clinicopathologic characteristics and survival condi-tions of BLBCL patients. Probably, A20 mutation is a factor associated with a poor prognosis of DLBCL.

Relation between Caspase recruitment domain-containing membrane-associated guanylate kinase protein 1, clinicopathological features of gastrointestinal lymphoma also and Helicobacter pylori infection / 中华消化杂志

Objective To investigate the relation between the expression of Caspase recruitment domain-containing membrane-associated guanylate kinase protein 1 (CARMA1),clinicopathological features of gastrointestinal mucosa-associated lymphoid tissue (MALT) lymphoma,diffuse large B cell lymphoma (DLBCL),and Helicobacter pylori (H.pylori) infection.Methods From January 1999 to March 2011,34 patients with DLBCL and 20 patients with MALT lymphoma were selected,and at same period 21 cases with reactive hyperplasia of gastrointestinal lymphoid tissue were enrolled in as control.The expression of CARMA1,Ki-67 and cytotoxin-associated gene A (CagA) at protein level were examined by immunohistochemistry.The relative expression quantity of CARMA1 mRNA was detected by real-time polymerase chain reaction (PCR).The condition of H.pylori infection in 25 gastric lymphoma and 10 controls was detected by methylene boric acid and blue staining or semi-nested PCR.Chi-square test was used for counting data analysis,t test for measurement data.Multivariate COX regression analysis was implemented for survival analysis.Survival curve was drawn by Kaplan-Meier method and analyzed by Log-rank test.Results The relative expression quantity of CARMA1 mRNA of 28 patients with DLBLC (3.073±1.846) was higher than that of 14 patients with MALT lymphoma,and the difference was statistically significant (F 0.975,P＜ 0.05).The positive rate of CARMA1 expression at protein level of gastrointestinal lymphoma group (75.9 ％,41/54) was higher than that of control group (47.6％,10/21),and the difference was statistically significant (x2 =5.568,P＜0.05),and the positive rate of CARMA1 expression of MALT lymphoma group (11/20) was lower than that of DLBCL group (88.2％,30/34),and the difference was statistically significant (x2 =5.900,P＜0.05).Among 42 patients with gastrointestinal lymphoma who received surgery,the relative expression quantity of CARMA1 mRNA in cases with high proliferation (2.885±1.837) was higher than that in cases with low proliferation.The expression of CARMA1 mRNA in the cases at advanced stage of the disease (4.416± 1.010) was higher than that in cases at early stage,and the difference was statistically significant (F=3.317 and 2.972,both P＜0.05).Among 54 patients with gastrointestinal lymphoma,the positive rate of CARMA1 expression at protein level of patients with high proliferation (88.6％,31/35) was higher that that of patients with low proliferation (10/19),and the difference was statistically significant (x22 ＝ 6.847,P＜0.05).There were no significant differences in relative expression quantity of CARMA1 mRNA and the positive rate of CARMA1 expression at protein level between 11 gastric lymphoma patients without H.pylori infection and 14 gastric lymphoma patients with H.pylori infection (both P＞0.05).The positive rate of CARMA1 expression at protein level of CagA positive and CagA negative H.pylori infected gastric lymphoma patients was 11/11 and 2/3.The expression of CARMA1 at protein level was correlated with the prognosis of gastrointestinal lymphoma (RR＝4.160,P＜0.05).In the 29 cases of patients with gastrointestinal MALT lymphoma and 18 cases of patients with DLBCL who were followed up,the survival situation of gastrointestinal lymphoma patients with CARMA1 positive expression rate over 50％ was worse than that of patients with the rate less than 50％,and the difference was statistically significant (x2=5.383 and 4.028,both P＜0.05).Conclusions CARMA1 might be involved in the pathogenesis and progression of MALT and DLBCL; and it might be a related factor of poor prognosis.There was no correlation between the expression of CARMA1 and H.pylori infection in these two lymphomas.

Study on optimal allocation of health resources in China based on equity and efficiency:strategy options and implemental paths / 中华医院管理杂志

Grounded on the review of the policies and outcomes of health resources allocation in China,the authors analyzed the allocation of health resources and its equity and efficiency,the comparison of and comment on domestic and foreign literatures.Based on such studies,they presented a strategic framework concerning optimal allocation of health resources in China with synergy of equity and efficiency,and developed an index system and an assessment system,as well as their optimal models and ways.

Inhibition of Emodin on LPS-induced Nitric Oxide Generation by Suppressing PLC-γ Phosphorylation in Rat Peritoneal Macrophages / 中草药·英文版

Objective To investigate the inhibitory mechanism of emodin on lipopolysaccharide(LPS)-induced nitric oxide(NO)generation in rat peritoneal macrophages.Methods NO production and iNOS expression were measured through nitrite assay and Western blotting assay,respectively.NF-kB activity and nuclei P65 expression were estimated by dual-luciferase and Western blotting assay,respectively.Intracellular free Ca2+([Ca2+]i)was detected using the ratiometric fluorescent calcium indicator dye,Fura-2,and a microspectrofluorometer.PLC-γphosporylation was analyzed by Western blotting assay.Results First,emodin was found playing active roles in suppressing LPS-induced NF-kB activation in rat peritoneal macrophages.Second,emodin down-regulated transient[Ca2*]i and could increase in NF-kB upstream signal.Finally,emodin suppressed phosphorylation of PLC-γ by LPS stimulation in the upstream of[Ca2+]i.Conclusion Suppression of PLC-γ phosphorylation is involved in emodin inhibiting NO generation by LPS stimulation in rat peritoneal macrophages.

Clinical Significance of MALT1 Gene Detection in Extra-nodal B Cell Lymphoma / 中国肿瘤临床

Objective: To investigate the relationship between MALT1 mRNA expression in extranodal B cell lymphoma and the pathogenesis of MALToma and DLBCL, and to explore the effect of MALT1 mRNA expression on the clinicopatho-serve the expression of Bcl-2 and Ki-67 in 106 samples of extranodal B cell lymphoma.The mRNA of MALT1 was detected by RT-PCR.Clinicopathological data were reviewed.Follow-up and statistical analysis were performed.Results: Expression of MALT1 mRNA was statistically different between the cases with lymph node metastasis and those without lymph node mestastasis, and between staging Ⅰ-Ⅱ cases and stage Ⅲ-Ⅳ cases, The expression of MALT1 mRNA in cases with posi-tive expression of MALToma, DLBCL, and Bcl-2 was higher than in those without expression of the three indices (P<0.05).However, no significant difference was found in MALT1 mRNA expression between cases with Ki-67≥30% and those with Ki-67<30% (P>0.05).MALT1 mRNA-positive cases showed a worse survival status than MALT1 mRNA-negative cases (P<0.05).Conclusion: Expression of MALT1 gene is different between MALToma and DLBCL.Inhibition of apoptosis by Bcl-2 overexpression caused by activation of NF-κB may lead to the pathogenesis of MALToma and DLBCL.The expression of MALT1 mRNA is associated with the types of lymphoma, involvement of lymph node, stage of lymphoma, and patient sur-vival.The expression of MALT1 gene may be associated with poor prognosis of MALToma and DLBCL.

Study on the options of medical institutions for residents in Tianjin city and influence factors / 中华医院管理杂志

Objective To probe into the variations of hospital admissions and rate of hospital bed occupancy of medical institutions in the recent ten years in Tianjin, for the purpose of formulating a regional health plan and optimizing regional health resources allocation. Methods The health statistics yearbooks of Tianjin from 1998 to 2008 and the four health service survey data from 1993 to 2008 were analyzed, with the descriptive statistics and ordinal logistic regression of SPSS 16. 0. Results From 1998 to 2008, the amplitude of hospital admissions and rate of hospital bed occupancy in community health service centers (stations) was the lowest, and the occupancy rate of hospital beds in community health service centers and rural hospitals was always lower than that in secondary and above hospitals. The influence factors of options of medical institutions for residents consist of hospital charges, gender,occupation, and regional classification. Conclusion Tianjin needs to strengthen its community health service centers , and improve service capacity of primary care of health organizations.

Research on Educational Cost Measurement of Higher Vocational Medical Education in Tianjin / 中华医学教育探索杂志

According to the characteristics of the higher vocational medical education,the paper discusses the higher vocational medical education cost measurement and the other questions.On the basis of analysis,it gives some solutions to it.