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.

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.

Objective To study the performance verification of lactate dehydrogenase(LDH) in the Johnson Vitros 5 .1 FS bio‐chemical analyzer .Methods According to CLSI instrumentation evaluation standard and referring to the validation scheme provided by the Johnson company ,the precision ,accuracy ,linear range of LDH ,maximum dilution degree ,biological reference range were verified .Results The LDH intra‐batch and inter‐batch precision experiments were≤3 .30% ;the accuracy experiment≤4 .00% ;the determination coefficient of the linear experiment was 0 .997 2 ;the LDH maximum dilution degree was 8 times with saline solution dilution;the biological reference range experiment verified that the reference range 313-618 U/L provided by the VITROS Meth‐odology Manual could be quote .Conclusion The performance verification of LDH detected by the Johnson Vitros 5 .1 FS biochemi‐cal analyzer basically conforms to the requirements of the quality objectives and manufacturer′s instructions ,and meets the needs of clinical test .

Objective:To investigate the molecular interaction between non-structural protein 3 serine protease of hepatitis C virus(HCV)and wild type P53,and to lay the basis for elucidating the mechanism of oncogenesis of hepatocellular carcinoma(HCC)after infection of HCV.Methods:The recombinant plasmids,pGAD424-NS3,pGAD424-NS315aa- and pGAD424-NS330aa-,were constructed and the interaction between NS3 serine protease and its cofactor NS4A,the interaction between wild type P53 and NS3 serine protease and its N-truncated mutants were dectected qualitatively and quantitatively in yeast two-hybrid system.Results:The results indicated that interaction existed not only between full-length NS3 serine protease and P53,but also between N-truncated mutants of NS3 serine protease and P53.Furthermore,the difference between enzyme activity unit(IU)of β-gal induced by these interactions was not significant(P>0.05).Conclusions:NS3 serine protease of hepatitis C virus and its N-truncated mutants can interact with wild type P53,and the region of NS3 serine protease involved in the interaction may be located in its C-terminal,but not in its N-terminal.

Objective To investigate clinical features of lower motor neuron lesion (LMNL) caused by the single level lower thoracic disc protrusion (LTDP),and to observe clinical outcomes of surgical treatment.Methods Between January 1997 and December 2009,17 patients with LMNL caused by single level LTDP underwent en bloc resection of the superior articular process,Cave-in 360° circumferential decompression and internal fixation in our hospital.MRI and CT scans were taken to confirm lesion levels:T10-11 in 4 patients of whom 3 had patellar clonus and ankle clonus,T11-12 in 5 patients of whom 4 had ankle clonus,and T12L1 in 8 patients who only had positive Babinski sign.The neurologic status was assessed using the Japanese Orthopaedic Association (JOA) scoring system.The muscle strength of the tibialis anterior was assessed using the Manual Muscle Test (MMT).Sagittal Cobb angle and cross-sectional area of the dural sac at the level of maximal compression in MRI were also observed.Results All patients were followed up for 22 to 76 months (average,48.6 months).The mean JOA score increased from preoperative 5.88±1.11 to 9.53±0.94 at final follow-up (t=16.143,P＜0.05).The muscle strength of the tibialis anterior recovered to more than grade 4 in all patients.Postoperative Cobb angle was unchanged compared with that before operation.MRI indicated that the cross-sectional area of the dural sac at the level of maximum compression increased from preoperative 35.8±7.3 mm2 to postoperative 132.9±6.5 mm2 (t=70.78,P＜0.05).Conclusion LMNL can be caused by LTDP.The eu bloc resection of the superior articular process,Cave-in 360° circumferential decompression and internal fixation can provide a satisfactory decompression effect and marked recovery of neurological function.

Objective To summarize our experience in the diagnosis and treatment of thyroid Hurthle cell neoplasms. Methods Clinical data of thyroid Hurthle cell neoplasm patients admitted from 1972 to 2003 were analyzed retrospectively. Results The main symptoms were thyroid solitary node or mass (37 cases) and multiple nodule (9 cases). 4 cases had cervical lymph node metastasis. With BUS、CT、ECT and FNAC,only 10 cases were diagnosed as HCNs preoperatively, 36 cases were misdiagnosed as nodular benign single or multiple node. Twenty-eight cases were diagnosed by FNAC and quick freezing pathology. Benign HCNs was treated by isolateral thyroidectomy and malignant HCNs by additional contralateral subtotal thyroidectomy. Eighteen cases were diagnosed by postoperative paraffin pathology, and retrival second operation was performed according to the benignity or malignancy of the thyroid lesion. All cases were followed-up for 2 to 10 years and doing well without recurrence. Conclusions We should realize thyroid Hurthle cell neoplasms fully, if HCNs is suspected intraoperatively, quick freezing pathology is helpful. With appropriate therapy, the prognosis is satisfactory.

Objective To investigate the current status about the application of alteplase (rt-PA) for intravenous thrombolysis in acute ischemic stroke patients,and clarify the relevant factors affecting patients'compliance of intravenous thrombolysis.Methods The acute ischemic stroke patients admitted in Department of Emergency,from January 2014 to December 2015 were recruited for study prospectively.After the patients with contraindications of thrombolysis were excluded,the eligible patients were divided into two groups,intravenous thrombolysis group (ITG) and non-intravenous thrombolysis group (NTG).Receiver operating characteristic curve (ROC) was used to determine the optimal cutoff point and the crucial NIHSS score of patients for decision on thrombolysis therapy.Results There were 230 patients with acute ischemic stroke occurred in the period of two years.Of 189 eligible patients,33 refused the intravenous thrombolysis treatment (NTG) whereas 156 willing to take the intravenous thrombolysis treatment (ITG).The intravenous thrombolysis rate of eligible ischemic stroke patients reached to Henan Provincial People's Hospital within the time window (4.5 hours) was 67.8％ without contraindications.The results of the single-factor analysis for the patients of the two groups displayed that the differences in factors including age,baseline NIHSS score,limb weakness,hemiplegic paralysis,dysphasia,as well as dizziness were significant between two groups (t =2.578,P =0.047;U =157.221,P =0.000;x2 =26.702,P =0.000;x2=9.069,P =0.003;x2 =7.381,P =0.007;x2 =28.636,P =0.000).The ROC analysis demonstrated the relationship between the baseline NIHSS score and the patients receiving intravenous thrombolysis.When NIHSS score ＜ 7,patients tended to refuse the treatment with intravenous thrombolysis (sensitivity 0.87,specificity 0.82).Among the patients receiving intravenous thrombolysis,the significant differences in intracranial hemorrhage rate,hospitalization mortality rate and 3-month mortality rate were not found between the patients with baseline NIHSS score≥7 and score ＜7 (1.9％ vs.3.9％,P =0.662;1.9％ vs.7.8％,P =0.168 and 3.8％ vs.11.7％,P =0.142,respectively).However a higher rate of favorable prognosis (3-month modified Rankin Scale score ≤ 1) was observed in thrombolysis patients (75.5％ vs.41.7％,P =0.000).Conclusions Factors such as age,baseline NIHSS score,limb weakness,hemiplegic paralysis,dysphasia,as well as dizziness are supposed to be associated with patients' compliance of intravenous thrombolysis.

NMDA-induced excitotoxicity cause severe neuronal damage including apoptosis and necrosis. The present study was aimed to evaluate the proportion of NMDA-induced apoptosis of rat cortical neurons and discover signal transduction mechanism. Caspase inhibitor and lactate dehydrogenase (LDH) assay were used to study the NMDA-induced apoptosis. To explore the involved signal pathways, the primary culture of rat cortical neurons were pretreated by the inhibitors of three MAPK pathways, extracellular signal-regulated kinase (ERK), c-Jun N-terminal kinase (JNK), and p38 MAPK. With 2 h of NMDA treatment, cellular apoptosis was measured by caspase-3 activity, terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick-end labeling (TUNEL) and Annexin V staining. The results showed that: (1) Caspase-dependent apoptosis accounted for 22.49% in NMDA-induced neuronal death; (2) Pretreatment with p38 MAPK inhibitor SB203580 (10 μmol/L) significantly decreased NMDA-mediated caspase-3 activity by 30.43% (P < 0.05). However, ERK inhibitor PD98059 (20 μmol/L) or JNK inhibitor SP600125 (20 μmol/L) did not influence caspase-3 activity; (3) Pretreatment with SB203580 significantly reduced the number of NMDA-induced TUNEL-positive cells by 33.10% (P < 0.05). PD98059 (20 μmol/L) or SP600125 (20 μmol/L) did not show obvious effect; (4) Pretreatment with SB203580 (10 μmol/L) significantly reduced the number of NMDA-induced early apoptotic neurons by 55.56% (P < 0.05). Also, SP600125 (20 μmol/L) significantly decreased the amount of late apoptotic/dead cells by 67.59% (P < 0.05). There was no effect of PD98059 (20 μmol/L). These results indicate that: (1) NMDA induces neuronal apoptosis besides necrosis; (2) p38 MAPK, but not JNK and ERK, is involved in NMDA-induced neuronal apoptosis, and inhibition of the apoptotic signaling pathway contributes to neuroprotection; (3) JNK activation might contribute to NMDA-induced neuronal necrosis rather than apoptosis.
Animals ; Anthracenes ; pharmacology ; Apoptosis ; Caspase 3 ; metabolism ; Cells, Cultured ; Extracellular Signal-Regulated MAP Kinases ; antagonists & inhibitors ; Imidazoles ; pharmacology ; JNK Mitogen-Activated Protein Kinases ; antagonists & inhibitors ; MAP Kinase Signaling System ; N-Methylaspartate ; pharmacology ; Neurons ; cytology ; Primary Cell Culture ; Pyridines ; pharmacology ; Rats ; p38 Mitogen-Activated Protein Kinases ; antagonists & inhibitors

OBJECTIVE Toprepareapolyclonalantibodyforspindlin1protein,anovelcancer related protein,and to provide the data for a better understanding of its functions and screening tu mor. METHODS Purifiedspindlin1proteinwasinjectedintorabbitstoproducethepolyclonalantiserumafter removing glutathione S-transferase (GST)from the fusion protein spindlin 1-GST that was expressed in Escherichia coli..The antiserum was purified through the Hitrap Protein A system,and the titer of spin-dlin 1 polyclonal antibody was detected by ELISA.The specificity of the polyclonal antibody was deter-minedbyWesternblottingandimmunohistochemistry.RESULTS Thetiterofspindlin1polyclonalanti-body was 1∶2000.Western blotting detection demonstrated that the spindlin 1 polyclonal antibody recog-nized myc-spindlin 1 reco mbinant fusion protein in HeLa cells transfected with pAdeasy-myc-spindlin 1 , which also corresponded with Myc.antibody.The HeLa cells were transfected with enhanced green fluo-rescence protein (EGFP)and spindlin 1 vector(pEGFP-C3-spindlin 1 ),which was confirmed by the in-dependent GFP fluorescence assay.The results of immunohistochemistry detection with the spindlin 1 polyclonal antibody suggested that spindlin 1 was mainly expressed in the nuclei of HeLa cells.More i m-portantly,in i mmunohistoche mical assays,the spindlin 1 antibody recognized nuclear spindlin 1 expres-sioninclinicalovariancancertissues.CONCLUSION Thespecificspindlin1polyclonalantibodyispre-pared,which may be used to detect cancer-related protein spindlin 1 in HeLa cells and ovarian cancer tissues.

Objective To investigate the iodine nutritional status of school children, lactating women and infants in iodine deficient areas of Baicheng and Wushi Counties in Xinjiang Autonomous Region. Methods According to the geographic location of east, south, west and north of county, 10 primary schools of 8 townships were selected. In each school, 10 children among each gender and age group from 8 to 10 years old were selected. A total of 300 school children were chosen. One hundred and four infants aged 0-2 years old and their mother were selected in 8 townships. Iodine content in edible salt at household level, the urinary iodine in school children and lactating women, total goiter rate(TGR) and the status of the intelligence quotient (1Q) of school children, the status of psychological development of infants were observed. Direct titration assay for testing the salt iodine, colorimetric ceric-arsenic assay and vitriolic ammonium assimilation were used for testing urinary iodine. The size of thyroid gland was measured by palpation. The Combined Raven Test for Chinese Rural was used to test the IQ. The psychological development of infants was tested by Danver Development Screening Test (DDST). Results The coverage rate of iodized salt at household level was 73.1% (123/182), however, the proportion of households using adequately iodized salt was 64.1% (118/182). The medium of urinary iodine in school children was 103.7 μ/L, with 47.8%(75/157) less than 100 μg/L and 28.0% (44/157) less than 50 μg/L; it was 123.0 μg/L in Baieheng County, with 44.4%(28/63) less than 100 μg/L and 33.3%(21/63) less than 50 μg/L; it was 100.3 μg/L in Wushi County, 50.0%(47/94) less than 100 μg/L and 24.5%(23/94) less than 50 μg/L. The medium of urinary iodine in locating women was 143.3 μg/L, it was 119.7 μg/L and 184.6 μg/L in Baicheng and Wushi Counties, respectively. The total rate of goiter in school children was 14.3%(43/300), it was 10.8%(13/120) and 16.6%(30/180) in Baicheng and Wushi Counties, respectively. The average IQ in school children was 80.6±11.6, it was 83.0±11.6 and 79.0±11.7 in Baicheng and Wushi Counties, respectively. The proportion of mental retardation in school children (IQ≤69) was 13.0% (39/300), it was 6.7% (8/120) and 17.2%(31/180) in Baicheng and Wushi Counties, respectively. In addition, the proportion of psychological development in infants being normal, suspicious and abnormal was 78.8%(82/104), 12.5% (13/104) and 8.7%(9/104), respectively. Conclusion This study confirms the fact that there is also existence of mental retardation in children and infants, caused by iodine deficiency.