BACKGROUND The connection between inflammatory bowel disease(IBD)and colorectal cancer(CRC)is well-established,as persistent intestinal inflammation plays a substantial role in both disorders.Cytokines may further inf...BACKGROUND The connection between inflammatory bowel disease(IBD)and colorectal cancer(CRC)is well-established,as persistent intestinal inflammation plays a substantial role in both disorders.Cytokines may further influence the inflammation and the carcinogenesis process.AIM To compare cytokine patterns of active IBD patients with early and advanced CRC.METHODS Choosing a panel of cytokines crucial for Th17/Treg differentiation and behavior,in colon specimens,as mRNA biomarkers,and their serum protein levels.RESULTS We found a significant difference between higher gene expression of FoxP3,TGFb1,IL-10,and IL-23,and approximately equal level of IL-6 in CRC patients in comparison with IBD patients.After stratification of CRC patients,we found a significant difference in FoxP3,IL-10,IL-23,and IL-17A mRNA in early cases compared to IBD,and IL-23 alone in advanced CRC.The protein levels of the cytokines were significantly higher in CRC patients compared to IBD patients.CONCLUSION Our findings showed that IL-6 upregulation is essential for both IBD and CRC development until the upregulation of other Th17/Treg related genes(TGFb1,IL-10,IL-23,and transcription factor FoxP3)is a crucial primarily for CRC development.The significantly upregulated IL-6 could be a potential drug target for IBD and prevention of CRC development as well.展开更多
BACKGROUND Cervical intraepithelial neoplasia(CIN)is an important precursor of cervical cancer.Early detection and treatment can reduce the incidence of cervical cancer.AIM To investigate the detection rate of human p...BACKGROUND Cervical intraepithelial neoplasia(CIN)is an important precursor of cervical cancer.Early detection and treatment can reduce the incidence of cervical cancer.AIM To investigate the detection rate of human papillomavirus(HPV)E6/E7 mRNA in cervical tissue of patients with different types of epithelial cell neoplasia(CIN)and its relationship with CIN progression and diagnosis.METHODS One hundred women with HPV infection detected by cervical exfoliation cytology between January 2022 and January 2023 were retrospectively selected.These patients were graded CIN based on colposcopy and cervical pathology.The positive expression rates of HPV E6/E7 mRNA and HPV[polymerase chain reaction(PCR)-reverse dot crossing]were compared among all groups.Patients with HPV E6/E7 mRNA expression in the grade 1 CIN group were followed up for 1 yr.The relationship between atypical squamous epithelium and high malignant epithelial neoplasia was investigated by univariate and multivariate analysis.RESULTS The diagnostic sensitivity,specificity,and sensitivity of PCR-reverse point hybrid ization technology for secondary CIN were 70.41%,70.66%,and 0.714,respectively.Sensitivity and specificity for secondary CIN were 752%and 7853%,respectively,the area under the curve value was 0.789.Logistic Multifactorial model analysis revealed that the HPV positive rates and the HPV E6/E7 mRNA positive rates were independent risk factors of CIN grade I(P<0.05).In CIN grade I patients with positive for HPV E6/E7 mRNA,in its orientation to grade CIN patients,in its orientation to grade CIN patients,at 69.2%,compared with patients negative for HPV E6/E7 mRNA(30.8%),significant difference(P<0.05).CONCLUSION HPV E6/E7 mRNA and HPV(PCR-reverse dot hybrid)positive expression have a close relationship with CINgrade disease progression and is an independent risk factor for high-grade CIN lesions.展开更多
High grade anal intraepithelial neoplasia due to human papilloma viral(HPV)infections is a precursor lesion for squamous cell carcinoma especially in high risk populations.Frequent examination and anal biopsies remain...High grade anal intraepithelial neoplasia due to human papilloma viral(HPV)infections is a precursor lesion for squamous cell carcinoma especially in high risk populations.Frequent examination and anal biopsies remain unpopular with patients;moreover they are also risk factors for chronic pain,scarring and sphincter injury.There is lack of uniform,surveillance methods and guidelines for anal HPV specifically the intervals between exam and biopsies.The aim of this editorial is to discuss the intervals for surveillance exam and biopsy,based on specific HPV related biomarkers?Currently there are no published randomized controlled trials documenting the effectiveness of anal screening and surveillance programs to reduce the incidence,morbidity and mortality of anal cancers.In contrast,the currently approved screening and surveillance methods available for HPV related cervical cancer includes cytology,HPV DNA test,P16 or combined P16/Ki-67 index and HPV E/6 and E/7 mRNA test.There are very few studies performed to determine the efficacy of these tests in HPV related anal precancerous lesions.The relevance of these biomarkers is discussed in this editorial.Longitudinal prospective research is needed to confirm the effectiveness of these molecular biomarkers that include high risk HPV serotyping,P16 immuno-histiochemistry and E6/E7 mRNA profiling on biopsies to elucidate and establish surveillance guidelines.展开更多
Background: The new 5G telecommunication technology has stirred concerns about potential negative effects on human health by radiofrequency electromagnetic fields. As to whether skin biology can be affected by 5G wave...Background: The new 5G telecommunication technology has stirred concerns about potential negative effects on human health by radiofrequency electromagnetic fields. As to whether skin biology can be affected by 5G waves has remained an unsolved challenge despite recent studies dealing with this issue. In particular, a strategy for rational design of an assay allowing to 1) reproducibly evaluate and decipher the 5G effects on skin as well as 2) test the potential protective effects of cosmetic active ingredients, has yet to be found. Here we describe an in vitro model of human normal keratinocytes irradiated by 5G waves and show their impact on two biomarkers of inflammatory stress, i.e. interleukin-1β (IL-1β) and reactive oxygen species (ROS) production. In addition, the capacity of a tannin-rich plant extract to protect against 5G impact is evaluated. Materials and Methods: In the first series of experiments, monolayers of human normal keratinocytes were irradiated or not (control) by 5G waves (3.5 MHz) in an anechoic chamber and were incubated at 37˚C for 24 hours. At the end of the incubation period, extracellular IL-1β and intracellular ROS were quantified using specific ELISA and colorimetric assays, respectively. In the second series of experiments, the effect of an overnight pre-incubation with increasing concentrations of a tannin-rich plant extract was evaluated. Additionally, we studied in a prospective way the expression of a set of 88 genes selected for their relevance to keratinocyte homeostasis, in relation to the 5G challenge as well as the protective effect of a tannin-rich plant extract. Results: 5G waves significantly increased IL-1β production by 48.4% (p β and ROS production. Finally, the expression of 47 genes was modified by 5G waves and/or by the tannin-rich plant extract. Conclusion: This is to our knowledge the first evaluation of the impact of 5G technology on inflammatory biomarkers of human normal skin cells. Here we provide an innovative and pertinent tool to screen for natural compounds with protective effects against 5G waves to develop cosmetic products shielding against the potentially deleterious effects of electromagnetic waves on human skin.展开更多
The reliable three dimensional (3-D) structure of the extracellular ligand-binding domain (V106-P322) of human interleukin-6 receptor (hlL-6R) has been constructed by means of computer-guided homology modeling techniq...The reliable three dimensional (3-D) structure of the extracellular ligand-binding domain (V106-P322) of human interleukin-6 receptor (hlL-6R) has been constructed by means of computer-guided homology modeling techniques using the crystal structure of the extracellular ligand-binding region (K52-L251) of human growth hormone receptor (hGHR) as templet. The space location of some key residues which influence the combination ability between the receptor and the ligand has been observed and the effects of point mutagenesis of the four conservative cysteine residues on the space conformation are analyzed. The results show that the space conformation of the side-chain carboxyl of E305 plays a key role in the ligand-binding ability. Furthermore, the space conformation of the side-chain carboxyl of E305 is very important for the electrostatic potential complementarity between hlL-6R and hlL-6 according to the docking method.展开更多
Background Human antigen R （HuR） is a ubiquitously expressed member of the ELAV family, and has relatively high cytoplasmic abundance in lung tissue regenerating after injury. In this study, we investigated whether ...Background Human antigen R （HuR） is a ubiquitously expressed member of the ELAV family, and has relatively high cytoplasmic abundance in lung tissue regenerating after injury. In this study, we investigated whether mitogen-activated protein kinase （MAPK）-activated protein kinase 2 （MK2） and HuR participate in the tumor necrosis factor （TNF）-induced expression of interleukin-6 （IL-6）. Methods Human pulmonary microvascular endothelial cells were treated with TNF following short interfering RNAmediated knockdown of MK2 or HuR. Cell supernatants were collected to detect the mRNA and protein expression of IL-6 at different time points, The expression and half-life of IL-6 mRNA were then determined in cells that had been treated with actinomycin D. Finally, after knockdown of MK2, the cytoplasmic expression of HuR protein was analyzed using Western blotting. Results MK2 or HuR knockdown decreased both the mRNA and protein expression of IL-6 in TNF-stimulated cells. In MK2 knockdown cells, the half-life of IL-6 mRNA was reduced to 36 minutes, compared with 67 minutes in the control group. In HuR knockdown cells, the half-life of IL-6 mRNA decreased from 62 minutes to 24 minutes. Further analysis revealed that knockdown of MK2 resulted in reduced HuR protein expression in the cytoplasm. Conclusions MK2 regulates the TNF-induced expression of IL-6 by influencing the cytoplasmic levels of HuR.展开更多
Objective: To explore the clinical significance of the quantitative detection of human papillomavirus(HPV) E6/E7 mRN A in triage of patients with atypical squamous cells of undetermined significance(ASC-US) and low-gr...Objective: To explore the clinical significance of the quantitative detection of human papillomavirus(HPV) E6/E7 mRN A in triage of patients with atypical squamous cells of undetermined significance(ASC-US) and low-grade squamous intraepithelial lesion(LSIL).Methods: A cross-sectional screening study was conducted among women who underwent outpatient gynecological screening at the Obstetrics and Gynecology Hospital of Fudan University from September 2015 to July 2016. A total of 500 patients from our hospital with ASC-US or LSIL based on cytology testing were subjected to HPV DNA and HPV E6/E7 mRNA quantitative analysis.Results: The specificity of the HPV E6/E7 mRNA test for detecting ≥high-grade squamous intraepithelial lesion(HSIL+) was statistically higher than that of the HPV DNA test(61.3% vs. 40.0%, P< 0.05), whereas there was no significant difference in the sensitivity of HPV E6/E7 mRNA test and HPV DNA test(90.0% vs. 95.0%, P > 0.05). The positive rates of HPV in the participants tested by HPV E6/E7 mRNA and HPV DNA were, respectively, 42.8%(214/500) and 62.8%(314/500), with statistical significance(P < 0.05).Conclusions: The HPV E6/E7 mRNA test was slightly less sensitivity than that of the HPV DNA test for diagnosing HSIL+ in patients with ASC-US and LSIL, but the difference was not significant, although the specificity of the former was significantly higher. HPV E6/E7 mRNA detection can effectively reduce overdiagnosis and overtreatment of patients with ASC-US and LSIL and has important clinical value in triage of patients with ASC-US and LSIL.展开更多
Based on the spatial conformations of human interleukin-6 (hlL-6) derived from nuclear magnetic resonance analysis and human interleukin-6 receptor (hlL-6R) modeled with homology modeling method using human growth hor...Based on the spatial conformations of human interleukin-6 (hlL-6) derived from nuclear magnetic resonance analysis and human interleukin-6 receptor (hlL-6R) modeled with homology modeling method using human growth hormone receptor as template, the interaction between hlL-6 and its receptor (hIL-6R) is studied with docking program according to the surface electrostatic potential analysis and spatial conformation complement. The stable region structure composed of hlL-6 and hlL-6R is obtained on the basis of molecular mechanism optimization and molecular dynamics simulation. The binding domain between hIL-6 and hIL-6R is predicted theoretically. Furthermore, the especial binding sites that influence the interaction between hlL-6 and hlL-6R are confirmed. The results lay a theoretical foundation for confirming the active regions of hlL-6 and designing novel antagonist with computer-guided techniques.展开更多
Objective To examine the effect of recombinant human transforming growth factor β1 (rhTGF β1) alone or recombinant human interleukin 6 (rhIL 6) alone or in combination on proliferation inhibition of the hum...Objective To examine the effect of recombinant human transforming growth factor β1 (rhTGF β1) alone or recombinant human interleukin 6 (rhIL 6) alone or in combination on proliferation inhibition of the human leukaemia cell line. Methods In the present study, using the human monoblastic cell line (U 937 ) and human promyelocytic cell line (HL 60 ) as an in vitro model, we analyzed the effect of two cytokins on proliferation inhibition with rate of 3H TdR incorporation, the cellular content of DNA, DNA indices, the cell cycle and the expression of c myc mRNA. Results With administration of rhTGF β 1 and rhIL 6, U 937 cell growth was inhibited and the rate of 3H TdR incorporation inhibition was increased. There was a decrease in the cellular content of DNA and DNA indices. And no change in the cell cycle was observed after administration of rhTGF β 1 or rhIL 6. However, there was an increase in G 0/G 1 phase cells and a decrease in G 2M+S phase cells after administration of combination of rhTGF β 1 and rhIL 6. It was also found that rhIL 6 could inhibit proliferative responses of HL 60 cells, meanwhile the inhibition could be enhanced by rhTGF β 1. The rate of 3H TdR incorporation inhibition rose up to 39.89%, and DNA index fell to 1.00 following induction by rhIL 6 plus rhTGF β 1. Furthermore, G 0/G 1 phase cells increased while G 2M+S cells decreased. Conclusions These results suggest that combination of rhTGF β 1 and rhIL 6 acted in synergy to inhibit proliferation of both U 937 and HL 60 cell lines. Molecular hybridization test show that rhTGF β 1 alone, rhIL 6 alone or rhTGF β 1 and rhIL 6 in combination can inhibit U 937 and HL 60 cells expression of c myc mRNA in a time and dose dependent manner. rhTGF β 1 and rhIL 6 in combination synergistically inhibited c myc expression, which may be one of the machanisms for the actions of the two cytokines.展开更多
In this study the three-dimensional (3-D) model of the ligand-binding domain (V106-P322) of human interleukin-6 receptor (hlL-6 R) was constructed by computer-guided ho-mology modeling technique using the crystal stru...In this study the three-dimensional (3-D) model of the ligand-binding domain (V106-P322) of human interleukin-6 receptor (hlL-6 R) was constructed by computer-guided ho-mology modeling technique using the crystal structure of the ligand-binding domain (K52-L251) of human growth hormone receptor (hGHR) as templet. Furthermore, the active binding region of the 3-D model of hlL-6R with the ligand (hlL-6) was predicted. In light of the structural characteristics of the active region, a hydrophobic pocket shielded by two hydrophilic residues (E115 and E505) of the region was identified by a combination of molecular modelling and the site-directed or double-site mutation of the twelve crucial residues in the ligand-binding domain of hIL-6R (V106-P322). We observed and analyzed the effects of these mutants on the spatial conformation of the pocket-like region of hlL-6 R. The results indicated that any site-directed mutation of the five Cys residues (four conservative Cys residues: Cyst 21, Cys132, Cys165,展开更多
Glioblastoma-multiforme(GBM), the most aggressive glial tumor, has a worldwide age-adjusted incidence ranging from 0.59-3.69/100000 persons. Despite current multimodal-treatment approach, median-survival time and prog...Glioblastoma-multiforme(GBM), the most aggressive glial tumor, has a worldwide age-adjusted incidence ranging from 0.59-3.69/100000 persons. Despite current multimodal-treatment approach, median-survival time and progression-free survival(PFS) remains short. Glioblastomas display a variety of molecular alterations, which necessitates determining which of these have a prognostic significance. This is a case of a 45-yearold patient who presented with progressive slurring of speech and features of raised intracranial pressure. Computed tomography(CT) scan revealed a large heterogeneously enhancing lesion in the left front-temporalperisylvian region with solid, cystic areas, suggestive of malignant glioma. Partial tumor-excision was followed by concurrent chemo-radiotherapy. Histopathologically, the tumor was astrocytoma grade-IV. Patient had an extended PFS of 12 mo, with an overall survival of 26 mo. Primary-GBM was confirmed using molecular markers and the immunophenotypic signature was defined by evaluating systemic expression of human telomerase reverse transcriptase, interleukin-6, neutrophil-lymphocyte ratio, tissue inhibitor of metalloproteinases-1, human chitinase-3-like-protein-1(YKL-40) and high mobility group-A1. Current findings suggest that this signature can identify worst outcomes, independent of clinical criteria.展开更多
Objective: To explore the effect of Guzhen recipe (固真方, GZR) on mRNA expression of interleukin-2 (IL-2) and interleukin-6 (IL-6) in senile rats as well as the relationship between dehydroepiandrosterone (DHEA) and ...Objective: To explore the effect of Guzhen recipe (固真方, GZR) on mRNA expression of interleukin-2 (IL-2) and interleukin-6 (IL-6) in senile rats as well as the relationship between dehydroepiandrosterone (DHEA) and its sulfate (DHEAS) and the immune function regulation of GZR. Methods: Northern Blot and reverse transcriptase-polymerase chain reaction (RT-PCR) were used to detect the mRNA expression of IL-2 and IL-6, meanwhile coating-antibody-counting method was used to detect DHEA(S) level in serum. Results: Compared with the young group, the splenocyte mRNA expression of IL-2 in senile rats was lowered and that of IL-6 was increased along with the decrease of serum DHEA(S) level. GZR, however, could improve the above indices in the senile rats. Conclusion: GZR might regulate the immune function of aged rats through increasing the serum level of DHEA(S).展开更多
基金Supported by the Medical University of Sofia,No.22.2012-2013Trakia University of Stara Zagora,No.1.2016 and No.2.2017.
文摘BACKGROUND The connection between inflammatory bowel disease(IBD)and colorectal cancer(CRC)is well-established,as persistent intestinal inflammation plays a substantial role in both disorders.Cytokines may further influence the inflammation and the carcinogenesis process.AIM To compare cytokine patterns of active IBD patients with early and advanced CRC.METHODS Choosing a panel of cytokines crucial for Th17/Treg differentiation and behavior,in colon specimens,as mRNA biomarkers,and their serum protein levels.RESULTS We found a significant difference between higher gene expression of FoxP3,TGFb1,IL-10,and IL-23,and approximately equal level of IL-6 in CRC patients in comparison with IBD patients.After stratification of CRC patients,we found a significant difference in FoxP3,IL-10,IL-23,and IL-17A mRNA in early cases compared to IBD,and IL-23 alone in advanced CRC.The protein levels of the cytokines were significantly higher in CRC patients compared to IBD patients.CONCLUSION Our findings showed that IL-6 upregulation is essential for both IBD and CRC development until the upregulation of other Th17/Treg related genes(TGFb1,IL-10,IL-23,and transcription factor FoxP3)is a crucial primarily for CRC development.The significantly upregulated IL-6 could be a potential drug target for IBD and prevention of CRC development as well.
基金Scientific Research Project of Hubei Provincial Health Commission,No.WJ2021M189。
文摘BACKGROUND Cervical intraepithelial neoplasia(CIN)is an important precursor of cervical cancer.Early detection and treatment can reduce the incidence of cervical cancer.AIM To investigate the detection rate of human papillomavirus(HPV)E6/E7 mRNA in cervical tissue of patients with different types of epithelial cell neoplasia(CIN)and its relationship with CIN progression and diagnosis.METHODS One hundred women with HPV infection detected by cervical exfoliation cytology between January 2022 and January 2023 were retrospectively selected.These patients were graded CIN based on colposcopy and cervical pathology.The positive expression rates of HPV E6/E7 mRNA and HPV[polymerase chain reaction(PCR)-reverse dot crossing]were compared among all groups.Patients with HPV E6/E7 mRNA expression in the grade 1 CIN group were followed up for 1 yr.The relationship between atypical squamous epithelium and high malignant epithelial neoplasia was investigated by univariate and multivariate analysis.RESULTS The diagnostic sensitivity,specificity,and sensitivity of PCR-reverse point hybrid ization technology for secondary CIN were 70.41%,70.66%,and 0.714,respectively.Sensitivity and specificity for secondary CIN were 752%and 7853%,respectively,the area under the curve value was 0.789.Logistic Multifactorial model analysis revealed that the HPV positive rates and the HPV E6/E7 mRNA positive rates were independent risk factors of CIN grade I(P<0.05).In CIN grade I patients with positive for HPV E6/E7 mRNA,in its orientation to grade CIN patients,in its orientation to grade CIN patients,at 69.2%,compared with patients negative for HPV E6/E7 mRNA(30.8%),significant difference(P<0.05).CONCLUSION HPV E6/E7 mRNA and HPV(PCR-reverse dot hybrid)positive expression have a close relationship with CINgrade disease progression and is an independent risk factor for high-grade CIN lesions.
文摘High grade anal intraepithelial neoplasia due to human papilloma viral(HPV)infections is a precursor lesion for squamous cell carcinoma especially in high risk populations.Frequent examination and anal biopsies remain unpopular with patients;moreover they are also risk factors for chronic pain,scarring and sphincter injury.There is lack of uniform,surveillance methods and guidelines for anal HPV specifically the intervals between exam and biopsies.The aim of this editorial is to discuss the intervals for surveillance exam and biopsy,based on specific HPV related biomarkers?Currently there are no published randomized controlled trials documenting the effectiveness of anal screening and surveillance programs to reduce the incidence,morbidity and mortality of anal cancers.In contrast,the currently approved screening and surveillance methods available for HPV related cervical cancer includes cytology,HPV DNA test,P16 or combined P16/Ki-67 index and HPV E/6 and E/7 mRNA test.There are very few studies performed to determine the efficacy of these tests in HPV related anal precancerous lesions.The relevance of these biomarkers is discussed in this editorial.Longitudinal prospective research is needed to confirm the effectiveness of these molecular biomarkers that include high risk HPV serotyping,P16 immuno-histiochemistry and E6/E7 mRNA profiling on biopsies to elucidate and establish surveillance guidelines.
文摘Background: The new 5G telecommunication technology has stirred concerns about potential negative effects on human health by radiofrequency electromagnetic fields. As to whether skin biology can be affected by 5G waves has remained an unsolved challenge despite recent studies dealing with this issue. In particular, a strategy for rational design of an assay allowing to 1) reproducibly evaluate and decipher the 5G effects on skin as well as 2) test the potential protective effects of cosmetic active ingredients, has yet to be found. Here we describe an in vitro model of human normal keratinocytes irradiated by 5G waves and show their impact on two biomarkers of inflammatory stress, i.e. interleukin-1β (IL-1β) and reactive oxygen species (ROS) production. In addition, the capacity of a tannin-rich plant extract to protect against 5G impact is evaluated. Materials and Methods: In the first series of experiments, monolayers of human normal keratinocytes were irradiated or not (control) by 5G waves (3.5 MHz) in an anechoic chamber and were incubated at 37˚C for 24 hours. At the end of the incubation period, extracellular IL-1β and intracellular ROS were quantified using specific ELISA and colorimetric assays, respectively. In the second series of experiments, the effect of an overnight pre-incubation with increasing concentrations of a tannin-rich plant extract was evaluated. Additionally, we studied in a prospective way the expression of a set of 88 genes selected for their relevance to keratinocyte homeostasis, in relation to the 5G challenge as well as the protective effect of a tannin-rich plant extract. Results: 5G waves significantly increased IL-1β production by 48.4% (p β and ROS production. Finally, the expression of 47 genes was modified by 5G waves and/or by the tannin-rich plant extract. Conclusion: This is to our knowledge the first evaluation of the impact of 5G technology on inflammatory biomarkers of human normal skin cells. Here we provide an innovative and pertinent tool to screen for natural compounds with protective effects against 5G waves to develop cosmetic products shielding against the potentially deleterious effects of electromagnetic waves on human skin.
文摘The reliable three dimensional (3-D) structure of the extracellular ligand-binding domain (V106-P322) of human interleukin-6 receptor (hlL-6R) has been constructed by means of computer-guided homology modeling techniques using the crystal structure of the extracellular ligand-binding region (K52-L251) of human growth hormone receptor (hGHR) as templet. The space location of some key residues which influence the combination ability between the receptor and the ligand has been observed and the effects of point mutagenesis of the four conservative cysteine residues on the space conformation are analyzed. The results show that the space conformation of the side-chain carboxyl of E305 plays a key role in the ligand-binding ability. Furthermore, the space conformation of the side-chain carboxyl of E305 is very important for the electrostatic potential complementarity between hlL-6R and hlL-6 according to the docking method.
基金This study was supported by the National Natural Science Foundation of China （No. 81270138）, the Natural Science Foundation of Jiangsu Province （No. BK2011657 and No. BK20130402）, and the Medical Technology Innovation Foundation of Nanjing Military Command （No. CWS 12J008）.
文摘Background Human antigen R （HuR） is a ubiquitously expressed member of the ELAV family, and has relatively high cytoplasmic abundance in lung tissue regenerating after injury. In this study, we investigated whether mitogen-activated protein kinase （MAPK）-activated protein kinase 2 （MK2） and HuR participate in the tumor necrosis factor （TNF）-induced expression of interleukin-6 （IL-6）. Methods Human pulmonary microvascular endothelial cells were treated with TNF following short interfering RNAmediated knockdown of MK2 or HuR. Cell supernatants were collected to detect the mRNA and protein expression of IL-6 at different time points, The expression and half-life of IL-6 mRNA were then determined in cells that had been treated with actinomycin D. Finally, after knockdown of MK2, the cytoplasmic expression of HuR protein was analyzed using Western blotting. Results MK2 or HuR knockdown decreased both the mRNA and protein expression of IL-6 in TNF-stimulated cells. In MK2 knockdown cells, the half-life of IL-6 mRNA was reduced to 36 minutes, compared with 67 minutes in the control group. In HuR knockdown cells, the half-life of IL-6 mRNA decreased from 62 minutes to 24 minutes. Further analysis revealed that knockdown of MK2 resulted in reduced HuR protein expression in the cytoplasm. Conclusions MK2 regulates the TNF-induced expression of IL-6 by influencing the cytoplasmic levels of HuR.
基金supported by the Shanghai Science and Technology Committee Project(No.16411950200).
文摘Objective: To explore the clinical significance of the quantitative detection of human papillomavirus(HPV) E6/E7 mRN A in triage of patients with atypical squamous cells of undetermined significance(ASC-US) and low-grade squamous intraepithelial lesion(LSIL).Methods: A cross-sectional screening study was conducted among women who underwent outpatient gynecological screening at the Obstetrics and Gynecology Hospital of Fudan University from September 2015 to July 2016. A total of 500 patients from our hospital with ASC-US or LSIL based on cytology testing were subjected to HPV DNA and HPV E6/E7 mRNA quantitative analysis.Results: The specificity of the HPV E6/E7 mRNA test for detecting ≥high-grade squamous intraepithelial lesion(HSIL+) was statistically higher than that of the HPV DNA test(61.3% vs. 40.0%, P< 0.05), whereas there was no significant difference in the sensitivity of HPV E6/E7 mRNA test and HPV DNA test(90.0% vs. 95.0%, P > 0.05). The positive rates of HPV in the participants tested by HPV E6/E7 mRNA and HPV DNA were, respectively, 42.8%(214/500) and 62.8%(314/500), with statistical significance(P < 0.05).Conclusions: The HPV E6/E7 mRNA test was slightly less sensitivity than that of the HPV DNA test for diagnosing HSIL+ in patients with ASC-US and LSIL, but the difference was not significant, although the specificity of the former was significantly higher. HPV E6/E7 mRNA detection can effectively reduce overdiagnosis and overtreatment of patients with ASC-US and LSIL and has important clinical value in triage of patients with ASC-US and LSIL.
文摘Based on the spatial conformations of human interleukin-6 (hlL-6) derived from nuclear magnetic resonance analysis and human interleukin-6 receptor (hlL-6R) modeled with homology modeling method using human growth hormone receptor as template, the interaction between hlL-6 and its receptor (hIL-6R) is studied with docking program according to the surface electrostatic potential analysis and spatial conformation complement. The stable region structure composed of hlL-6 and hlL-6R is obtained on the basis of molecular mechanism optimization and molecular dynamics simulation. The binding domain between hIL-6 and hIL-6R is predicted theoretically. Furthermore, the especial binding sites that influence the interaction between hlL-6 and hlL-6R are confirmed. The results lay a theoretical foundation for confirming the active regions of hlL-6 and designing novel antagonist with computer-guided techniques.
文摘Objective To examine the effect of recombinant human transforming growth factor β1 (rhTGF β1) alone or recombinant human interleukin 6 (rhIL 6) alone or in combination on proliferation inhibition of the human leukaemia cell line. Methods In the present study, using the human monoblastic cell line (U 937 ) and human promyelocytic cell line (HL 60 ) as an in vitro model, we analyzed the effect of two cytokins on proliferation inhibition with rate of 3H TdR incorporation, the cellular content of DNA, DNA indices, the cell cycle and the expression of c myc mRNA. Results With administration of rhTGF β 1 and rhIL 6, U 937 cell growth was inhibited and the rate of 3H TdR incorporation inhibition was increased. There was a decrease in the cellular content of DNA and DNA indices. And no change in the cell cycle was observed after administration of rhTGF β 1 or rhIL 6. However, there was an increase in G 0/G 1 phase cells and a decrease in G 2M+S phase cells after administration of combination of rhTGF β 1 and rhIL 6. It was also found that rhIL 6 could inhibit proliferative responses of HL 60 cells, meanwhile the inhibition could be enhanced by rhTGF β 1. The rate of 3H TdR incorporation inhibition rose up to 39.89%, and DNA index fell to 1.00 following induction by rhIL 6 plus rhTGF β 1. Furthermore, G 0/G 1 phase cells increased while G 2M+S cells decreased. Conclusions These results suggest that combination of rhTGF β 1 and rhIL 6 acted in synergy to inhibit proliferation of both U 937 and HL 60 cell lines. Molecular hybridization test show that rhTGF β 1 alone, rhIL 6 alone or rhTGF β 1 and rhIL 6 in combination can inhibit U 937 and HL 60 cells expression of c myc mRNA in a time and dose dependent manner. rhTGF β 1 and rhIL 6 in combination synergistically inhibited c myc expression, which may be one of the machanisms for the actions of the two cytokines.
文摘In this study the three-dimensional (3-D) model of the ligand-binding domain (V106-P322) of human interleukin-6 receptor (hlL-6 R) was constructed by computer-guided ho-mology modeling technique using the crystal structure of the ligand-binding domain (K52-L251) of human growth hormone receptor (hGHR) as templet. Furthermore, the active binding region of the 3-D model of hlL-6R with the ligand (hlL-6) was predicted. In light of the structural characteristics of the active region, a hydrophobic pocket shielded by two hydrophilic residues (E115 and E505) of the region was identified by a combination of molecular modelling and the site-directed or double-site mutation of the twelve crucial residues in the ligand-binding domain of hIL-6R (V106-P322). We observed and analyzed the effects of these mutants on the spatial conformation of the pocket-like region of hlL-6 R. The results indicated that any site-directed mutation of the five Cys residues (four conservative Cys residues: Cyst 21, Cys132, Cys165,
基金Supported by M.P.Biotech Council,M.P.for financial assistance and BMHRC for infrastructural facilities,No.249
文摘Glioblastoma-multiforme(GBM), the most aggressive glial tumor, has a worldwide age-adjusted incidence ranging from 0.59-3.69/100000 persons. Despite current multimodal-treatment approach, median-survival time and progression-free survival(PFS) remains short. Glioblastomas display a variety of molecular alterations, which necessitates determining which of these have a prognostic significance. This is a case of a 45-yearold patient who presented with progressive slurring of speech and features of raised intracranial pressure. Computed tomography(CT) scan revealed a large heterogeneously enhancing lesion in the left front-temporalperisylvian region with solid, cystic areas, suggestive of malignant glioma. Partial tumor-excision was followed by concurrent chemo-radiotherapy. Histopathologically, the tumor was astrocytoma grade-IV. Patient had an extended PFS of 12 mo, with an overall survival of 26 mo. Primary-GBM was confirmed using molecular markers and the immunophenotypic signature was defined by evaluating systemic expression of human telomerase reverse transcriptase, interleukin-6, neutrophil-lymphocyte ratio, tissue inhibitor of metalloproteinases-1, human chitinase-3-like-protein-1(YKL-40) and high mobility group-A1. Current findings suggest that this signature can identify worst outcomes, independent of clinical criteria.
文摘Objective: To explore the effect of Guzhen recipe (固真方, GZR) on mRNA expression of interleukin-2 (IL-2) and interleukin-6 (IL-6) in senile rats as well as the relationship between dehydroepiandrosterone (DHEA) and its sulfate (DHEAS) and the immune function regulation of GZR. Methods: Northern Blot and reverse transcriptase-polymerase chain reaction (RT-PCR) were used to detect the mRNA expression of IL-2 and IL-6, meanwhile coating-antibody-counting method was used to detect DHEA(S) level in serum. Results: Compared with the young group, the splenocyte mRNA expression of IL-2 in senile rats was lowered and that of IL-6 was increased along with the decrease of serum DHEA(S) level. GZR, however, could improve the above indices in the senile rats. Conclusion: GZR might regulate the immune function of aged rats through increasing the serum level of DHEA(S).
