AIM:This study aimed to establish a model of functional impairment in INS-1 cells(rat insulino-ma cells)induced by prolonged exposure to a low level of interleukin-1β(IL-1β),and to investigate the underlying mech-anisms.METHODS:INS-1 cells were treated with different concentrations of IL-1β for 10 d.The cells were divided into control,0.05 μg/L IL-1β,0.1 μg/L IL-1β,and 0.2 μg/L IL-1β groups(n=3).Flow cytometry was employed to assess cell apoptosis,facilitating the identification of IL-1β concentrations that did not significantly induce apoptosis.Subse-quently,INS-1 cells were divided into control and IL-1β groups(n=3)and treated with 0.1 μg/L IL-1β for 10 days.The intracellular insulin content and glucose-stimulated insulin secretion level were detected by immunofluorescence and en-zyme-linked immunosorbent assay(ELISA).Differential proteins and associated signaling pathways were screened using 4D label-free proteomics technology.Moreover,fluorescent probes were used to detect glucose-stimulated calcium ion in-flux,and Western blot was conducted to verify the expression changes of relevant proteins in the nuclear factor kappa-B(NF-κB)signaling pathway and calcium signaling pathway.RESULTS:Treatment with 0.1 μg/L IL-1β for 10 days ex-hibited minimal impact on the survival of INS-1 cells but significantly reduced the intracellular insulin content and glucose-stimulated insulin secretion(P<0.01).Therefore,0.1 μg/L IL-1β was chosen for in vitro induction of functional impair-ment in INS-1 cells.Proteomic analysis showed no significant differences in the expression of apoptosis-related factors,such as caspase-3,B-cell lymphoma-2(Bcl-2),Bcl-2-associated X protein(Bax),and Bcl-2-associated death promoter(Bad),compared with the control(P>0.05).However,the expression of insulin,v-maf avian musculoaponeurotic fibro-sarcoma oncogene family protein A(Mafa),calcium/calmodulin-dependent protein kinase II(CaMKII),inositol 1,4,5-triphosphate receptor(IP3R),and NF-kappa-B inhibitor alpha(IκBα)was significantly reduced(P<0.01),while the protein expression levels of NF-κB and inducible nitric oxide synthase(iNOS)were significantly increased(P<0.05,P<0.01).Fluorescence probe results showed that,compared to the control,the calcium ion influx induced by glucose stimu-lation in the IL-1β group was significantly reduced(P<0.01).Western blot results demonstrated that the expression lev-els of IP3R,CaMKII,and IκBα in the IL-1β group were significantly reduced(P<0.05,P<0.01),while the expression levels of NF-κB and iNOS were significantly increased(P<0.01).CONCLUSION:A model of functional impairment in rat INS-1 cells induced by prolonged exposure to low levels of IL-1β has been successfully established,and it has been pre-liminarily confirmed that this impairment may be related to the activation of the NF-κB signaling pathway and dysfunction of the calcium signaling pathway.

AIM:This study aimed to establish a model of functional impairment in INS-1 cells(rat insulino-ma cells)induced by prolonged exposure to a low level of interleukin-1β(IL-1β),and to investigate the underlying mech-anisms.METHODS:INS-1 cells were treated with different concentrations of IL-1β for 10 d.The cells were divided into control,0.05 μg/L IL-1β,0.1 μg/L IL-1β,and 0.2 μg/L IL-1β groups(n=3).Flow cytometry was employed to assess cell apoptosis,facilitating the identification of IL-1β concentrations that did not significantly induce apoptosis.Subse-quently,INS-1 cells were divided into control and IL-1β groups(n=3)and treated with 0.1 μg/L IL-1β for 10 days.The intracellular insulin content and glucose-stimulated insulin secretion level were detected by immunofluorescence and en-zyme-linked immunosorbent assay(ELISA).Differential proteins and associated signaling pathways were screened using 4D label-free proteomics technology.Moreover,fluorescent probes were used to detect glucose-stimulated calcium ion in-flux,and Western blot was conducted to verify the expression changes of relevant proteins in the nuclear factor kappa-B(NF-κB)signaling pathway and calcium signaling pathway.RESULTS:Treatment with 0.1 μg/L IL-1β for 10 days ex-hibited minimal impact on the survival of INS-1 cells but significantly reduced the intracellular insulin content and glucose-stimulated insulin secretion(P<0.01).Therefore,0.1 μg/L IL-1β was chosen for in vitro induction of functional impair-ment in INS-1 cells.Proteomic analysis showed no significant differences in the expression of apoptosis-related factors,such as caspase-3,B-cell lymphoma-2(Bcl-2),Bcl-2-associated X protein(Bax),and Bcl-2-associated death promoter(Bad),compared with the control(P>0.05).However,the expression of insulin,v-maf avian musculoaponeurotic fibro-sarcoma oncogene family protein A(Mafa),calcium/calmodulin-dependent protein kinase II(CaMKII),inositol 1,4,5-triphosphate receptor(IP3R),and NF-kappa-B inhibitor alpha(IκBα)was significantly reduced(P<0.01),while the protein expression levels of NF-κB and inducible nitric oxide synthase(iNOS)were significantly increased(P<0.05,P<0.01).Fluorescence probe results showed that,compared to the control,the calcium ion influx induced by glucose stimu-lation in the IL-1β group was significantly reduced(P<0.01).Western blot results demonstrated that the expression lev-els of IP3R,CaMKII,and IκBα in the IL-1β group were significantly reduced(P<0.05,P<0.01),while the expression levels of NF-κB and iNOS were significantly increased(P<0.01).CONCLUSION:A model of functional impairment in rat INS-1 cells induced by prolonged exposure to low levels of IL-1β has been successfully established,and it has been pre-liminarily confirmed that this impairment may be related to the activation of the NF-κB signaling pathway and dysfunction of the calcium signaling pathway.

Clinical microbiology examination technology experiment is an important part of clinical microbiology examination technology teaching. In the experimental teaching of clinical microbiology examination technology, the virtual simulation technology was combined with traditional teaching to give full play to the advantages of the virtual experimental platform. As to experimental projects that couldn't be carried out in traditional teaching and some important experimental projects, students could learn on the virtual experimental platform, and after learning, they would participate in the corresponding assessment. The perfect combination of the two can solve the problem of high experimental cost and limited experimental content in the current experimental class, make up for the shortcomings of traditional teaching, realize the sharing of teaching resources. Besides, it can strengthen the students' experimental operation skills and enhance the interest of learning for cultivation of application-oriented medical talents.

Objective To investigate the differences of histopathological diagnosis between the endoscopic mucosal resection (EMR) specimens and the biopsy specimens,and to evaluate the value and the limitation of EMR in diagnosis of early esophageal cancers and its precursor lesions.Methods A retrospective analysis on 217 lesions with early esophageal cancers or the precursor lesions treated by EMR was performed.The differences between pathological diagnoses of biopsy and EMR were compared.Results Compared with pathologic diagnosis after EMR,the yield of biopsy consisted of 41.9％ (91/217) as under-diagnosed,15.7％ (34/217) as over-diagnosed,and 42.4％ (92/217) as consistent.EMR diagnosis also explicated the differentiation,the grade,the invasive depth and the lympho-vascular infiltration of the lesions.Conclusion The endoscopic biopsy diagnosis is limited for the pathological diagnosis of early esophageal cancer and precancerous lesions,while the EMR sample can provide objective diagnosis and provide the guideline for the further treatment.

Objective To evaluate the efficiency of BIOMED-2 system in detecting IGH gene clonal rearrangement and application in diagnosis of extranodal marginal zone B-cell lymphoma of mucosa-associated lymphoid tissue (MALT) lymphoma. Methods Forty-five cases were collected, including 36 MALT lymphomas from different organs, 3 extranodal lymphoid tissue proliferative lesions and 6 severe gastritis associated with H pylori. DNA was extracted from the formalin fixed paraffin embedded blocks of these cases and the quality of DNA was assessed using the BIOMED-2 specimen mixed control primers. IGH gene clonal rearrangement was detected using IGH VH-JH primers. The sensitivity and specificity of BIOMED-2 PCR were analyzed.Results Adequate DNA (≥ 300 bp) was obtained in 31 of 45 samples (including 22 MALT lymphomas, 3 lymphoid tissue proliferative lesions and 6 severe gastritis), and the DNA from the other 14 samples was degraded seriously. 16 of 22 MALT lymphomas were positive of IGH clonal rearrangement with the sensitivity of 72.7 ％.In contrast,none of 6 severe gastritis was positive with the specificity of 100 ％ IGH and clonal rearrangement were detected in one of 3 lymphoid tissue proliferative lesions. Conclusion BIOMED-2 assay is an effient and reliable method for diagnosis and differential diagnosis of MALT lymphoma which is important for clinical practical value.

OBJECTIVE: To evaluate the clinic manifestation, pathologic behavior, therapy and prognosis of rare aggressive fibromatosis in the head and neck. METHOD: Two cases of aggressive fibromatosis were analyzed and relevant literatures were reviewed. RESULT: Aggressive fibromatosis was characterized as infiltrative, locally aggressive and tended to recur after surgical resection. Pathology showed fibroblastic monoclonal proliferation. Fibromatosis was composed of well-differentiated fibroblasts and myofibroblasts, lacking cytological features of malignancy and scanty or absent mitotic activity. Complete surgical excision of aggressive fibromatosis was considered to be the only effective method of cure by most authorities. Chemotherapy and radiotherapy can be used together with surgery in recurrence or unsatisfactory surgical margin. In our study, one patient recurred after the first operation, and after another operation, the patient did not recur after 6 months follow up, and the other one did not recur after 6 months follow up. CONCLUSION The diagnosis of aggressive fibromatosis depended on pathological examination. Radical removal was an important way to reduce recurrence rate. Radiation therapy and chemotherapy can be used as adjuvant therapy in patients with recurrent or unresectable or inoperable disease.
Adolescent ; Female ; Fibromatosis, Aggressive ; diagnosis ; surgery ; Head and Neck Neoplasms ; diagnosis ; surgery ; Humans ; Middle Aged

Objective To assess the role of promyelocytic leukemia protein (PML)and P53 in the progression of esophageal squamous cell carcinoma(ESCC)and its precursor lesions. Methods Different expression patterns of PML and P53 of 241 cases of ESCC combined with adjacent precursors were analyzed by tissue array and immunohistochemistry and correlated with clinicopathological parameters. Results In ESCC and its precursor lesions, PML and P53 displayed positive or strong positive, while in normal esophageal epithelia, these proteins showednegative or stained positive only in parabasal cell layer. The expression level of PML was correlated with the depth of invasion of esophageal carcinomas (X2=29.461,P<0.001),lymph metastasis status(X2=15.226,P<0.001)and pTNMs(x2=26.956,P

Objective To clarify the MUM1/IRF4 expression in follicular lymphoma (FL) and its clinical and pathological significance. Methods Ninety-six cases FL were immunostained with MUM1,CD10,bcl-2,bcl-6 and Ki-67 antibodies. The results were compared with their clinical and pathological features. Results The overall MUM1 expression rate in FL was 59.2 % (58/96),including 36.2 % (19/51) grade 1 or 2 and 86.4 %(39/45) grade 3 cases (x2 =24.406,P <0.001). 68.9 % cases with diffuse area were MUM1 positive (x2 =8.161,P =0.004). MUM 1 and CD10 expression had inverse correlation,83.3 % CD10 negative cases were MUM1 positive (x2= 12.649,P<0.001). The mitosis rate and Ki-67 label index were statistically higher in MUM1 positive cases than in negative cases (t = -3.852 & -4.610,respectively,P <0.001). Conclusion MUM1 can be used as a biomarker to divide FL into different malignancies. The MUM1 positive FL may be the feature of high grade non germinal center B cell malignant lymphoma.

Objective To analyze the clinical results and prognostic factors of patients with early-stage primary gastric mucosa-associated lymphoid tissue(MALT) lymphoma. Methods Seventy-seven pa-tients with primary gastric MALT lymphoma treated from 1985 to 2006 were retrospectively analyzed. All pa-tients were pathologically confirmed as MALT lymphoma in stage Ⅰ ,Ⅱ and ⅡE (by modified Blackedge staging system). Thirty-seven patients had stage Ⅰ disease,23 stage Ⅱ and 17 stage ⅡE. Sixty patients un-derwent surgical resection and 17 received non-surgical treatment. Survival rates were calculated by the Kap-lan-Meier analysis with the Logrank test. Results With a median follow up of 57 months for the surviving patients(ranging from 1 to 198 months for all patients), the 5-year overall survival rate, disease-free survival rate,loco-regional control rate and distant metastasis free survival rate were 74% ,70% ,76% and 87% ,re-spectively. In univariate analysis, clinical stage was significantly associated with overall survival. Patients with stage Ⅰ or Ⅱ disease had a better overall survival than those with stage ⅡE (P = 0.01). Tumor size and surgical resection were significantly associated with disease-free survival. Patients with primary tumor 8 cm or less in diameter had better disease-free survival than those with primary tumor more than 8 cm in diameter(P =0.03). Patients who underwent complete resection had better disease-free survival than those who under-went incomplete resection or no surgery (P =0.02). Clinical stage, tumor size and surgical resection were significantly associated with loco-regional control. Patients with stage Ⅰ or Ⅱ disease had better loco-regional control than those with stage ⅡE (P = 0. 03). Patients with primary tumor 8 cm or less in diameter had better loco-regional control than those with primary tumor more than 8 cm in diameter(P =0.01). Patients who un-derwent complete resection had better loco-regional control than those who underwent incomplete resection or no surgery(P=0.03). Patients with stage Ⅰ and Ⅱ disease treated with surgery had more local recurrence, and patients treated without surgery tended to recur systematically. Patients with stage ⅡE disease tended to recur locally in spite of surgery or not. Conclusions The efficacy of surgical and non-surgical treatment for primary gastric MALT lymphoma are similar. Surgical resection is no longer a necessary approach in the primary treatment. Clinical stage is an important prognostic factor for primary gastric MALT lymphoma.

Objective To evaluate the efficacy of endoscopic ultrasound guided fine-needle aspiration (EUS-FNA) in diagnosis of enlarged mediastinal lymph nodes (LNs), mediastinal occupying lesion of unknown origin, as well as in N-staging for lung cancer. Methods EUS-FNA was performed via esophagus with a 22-gange needle in 61 patients, followed by pathological and cytological examinations. Results The positive diagnosis rate of EUS-FNA was 93.4% (57/61), and the cytological and pathological diagnostic accuracy were 85.2% (52/61) and 83.6% (51/61), respectively. Of 61 patients, 26 were suspected as having lung cancer with mediastinal lymph nodes metastasis, but the bronchoscopy failed to confirm the diag-nosis. EUS-FNA diagnosed lung cancer in 21 and benign lesion in 5. Of 22 patients with mediastinal occupying lesions of unknown origin, 19 (86.4%) were diagnosed by EUS-FNA. Of 7 patients with malignant tumor history and enlarged mediastinal lymph nodes, EUS-FNA confirmed mediastinal metastasis in 6 (85.7%). Six cases of lung cancer with suspected mediastinal lymph nodes metastasis were confirmed by EUS-FNA and the corresponding therapy regimen was modified. No complications related to EUS-FNA procedure occurred. Conclusion EUS-FNA is a safe and effective method for diagnosis of enlarged medistinal LNs, mediastinal lesion of unkown origin and N-stage of lung cancer.