Objective To investigate the effect of Bcl-2 gene knockdown on apotosis , proliferation and drug sensitivity of gastric carcinoma HGC-27 cells. Methods The HGC-27 cells were divided into five groups:the untreated control group , the control siRNA group , the specific siRNA targeting Bcl-2 gene group , 5-FU treated group and the combination group (Bcl-2 siRNA and 5-FU treatment). Then flow cytometry and MTT assays were performed to detect the apoptosis and proliferation of HGC-27 cells. The cysteine protease activityand Cytochrome C release level were tested by ELISA method. Results Bcl-2 knockdown enhanced apoptosis and inhibited proliferation of HGC-27 cells. Comparedwith the 5-FU-treated group , the cell apoptosis level, activities of Caspase-3 and Caspase-9, plasma Cytochrome C were significantly increased in the combination group(P <0.01). Conclusion Bcl-2 gene knockdown induced apoptosis, inhibited cell proliferation and enhanced the drug sensitivity of 5-FU in gastric cancer cells , which might be considered as a potential therapeutic strategy forgastric carcinoma.

Objective To prepare the preparation of ginkgolic acids gel and to study its antibacterial activity in vitro.Methods The gel was prepared with ginkgolic acids as main component and with Carbopol 940 as base material.The content of ginkgolic acids in the gel was determined by high performance liquid chromatography and the minimum inhibitory concentration (MIC) of the gel against staphylocococus aureus and propionibacteria was measured in vitro by means of liquid-doubling dilution.Results The ginkgolic acids gel has inhibitory effects on staphylocococus aureus and propionibacteria to various extents.And the MIC was 31.756 mg?mL-1 for staphylocococus aureus and 0.992 mg?mL-1 for propionibacteria.Conclusion The preparation of the gel is feasiblye and the the properties of the gel were stable.The ginkgolic acids gel possesses good antibacterial effects on staphylocococus aureus and propionibacteria.

BACKGROUND: Allogeneic hematopoietic stem cell transplantation(allo-HSCT)is an effective method for treating multiple malignant hematological diseases and hereditary diseases.However,systematic internal organs disorders,especially pulmonary complications,are commonly following allo-HSCT,How to correctly diagnose and treat the coexistence of pulmonary infectious complications and pulmonary noninfectious complications has great importance.OBJECTIVE: To report a case suffered from pulmonary Aspergillus infection coexisted with obliterative bronchiolitis at 1 year following allo-HSCT,and to discuss the prevention,clinical manifestation and treating method by reviewing related literature.METHODS: At 373 days after allo-HSCT,the patient developed fever,dry cough,shortness of breath and dyspnea on exertion A high-resolution computed tomography of chest demonstrated that there were alveolar infiltrating in the upper,middle and lower lobe of the right lung,and the focus of infection was performed further biopsy.RESULTS AND CONCLUSION: The histopathological examination of the sample showed alveolus dilatation,epithelial cells hyperplasia,fibrinous obliteration in alveolar space and peribronchiolar lymphocytes inflammation,which were CD3(+),CD45RO(+),CD20(-),CD79a(-),MPO(-),CD34(-).Aspergillus fumigatus could be seen in the cultured biopsied tissue specimen.Pulmonary function test showed that,air flow obstruction with reduction of forced expiratory volume in one second was 59.27%.The patient was diagnosed as invasive pulmonary Aspergillus infection combined with bronchiolitis obliterans and was treated by caspofungin combined with intravenous voriconazole for invasive aspergillosis,methylprednisolone,azathioprine,intravenous immunoglobulin and azithromycin for bronchiolitis obliterans.At 40 days after treatment,the CT examination showed the focus was absorbed completely.

Objective To compare the diagnostic value of shear wave elastography and 18F-FDG PET/CT imaging in diagnosis of thyroid microcarcinoma.Methods Shear wave elastography and 18F-FDG PET/CT data of patients with thyroid nodules confirmed by pathology were analyzed retrospectively.The sensitivity,specificity,accuracy,positive and negative predictive value of the two methods in diagnosis of malignant thyroid micronodules were compared.Results In all of the 48 micronodules,37 were malignant,11 were benign.The sensitivity,specificity,accuracy,positive predictive value and negative predictive value of shear wave elastography and 18F-FDG PET/CT were 91.89％ (34/37),81.82％ (9/11),89.58％ (43/48),94.44％ (34/36),75.00％ (9/12) and 29.73％ (11/37),72.73％ (8/11),39.58％ (19/48),78.57％ (11/ 14),23.53％ (8/34),respectively.The sensitivity,accuracy and negative predictive value between the two methods had statistical difference (all P＜0.05),and the specificity and positive predictive value between the two methods had no statistical difference (both P＞0.05).Conclusion Compared to 18F FDG PET/CT imaging,the diagnositic value of shear wave elastography in diagnosis of thyroid microcarcinoma has obvious advantages.

Objective To investigate the clinicopathological significances of LDHA/mutant p53 co-expres-sion in gliomas. Methods According to the 2016 WHO CNS,archived 68 gliomas were collected and analyzed retrospectively. The co-expression of LDHA/mutant p53 was detected by immunohistochemical staining. Results High expression of LDHA alone was always found in high grade gliomas(48.5%). Mutant p53 high expression was usually observed in glioblastomas (26.5%). There was a close relationship between co-expression of LDHA/mutant p53 in glioblastoma(27.9%,P = 0.005),or gliomas with high histological grading(27.9%,P = 0.002). Conclusions Co-expression of LDHA/mutant p53 in tumor cells might be a specific immunohistochemical pheno-type of gliomas,and may help for distinguishing glioblastoma and other high grade gliomas from low grade gliomas.

Objective To investigate the potential pathogenesis of Focal cortical dysplasia (FCD), we performed cDNA microarray analysis to obtain gene expression profile of FCD. Methods Three FCD samples and three normal controls were enrolled. Total RNA of the brain tissues were extracted. The difference gene expressions between FCD group and control group was detected using Affymetrix gene chip. The up and down-regulated genes were confirmed by Real-time PCR. Further, the related signal pathways involved in the pathogenic mechanisms of FCD were predicted by bioinformatics. Result In FCD, two up-regulated genes C21orF2 and AU152162 and 5 down-regulated genes ENPP2, ANLN, IP6K3, UGT8, and AZGP were found. Compared the FCD samples with the normal controls , there were significantly different in all down-regulated genes (P < 0.05), while the up-regulated genes were not (P > 0.05). Using bioinformatics analysis, the ENPP2 , UGT8 , and AZGP1 protein which located in the cell membrane or secreted into the extracellular matrix may be involved in the formation of the myelin sheath and the development of the nervous system by the lipid metabolism and LPA signaling pathway. Conclusion ENPP2, UGT8 and AZGP1 may be involved in pathogenesis of FCD through the process of myelin sheath formation and LPA signal pathway , which warrants further study to know their roles in the pathogenesis of FCD.

OBJECTIVETo study the effect of hepatitis C virus nonstructural protein NS(3) (HCV NS3) on telomerase activity and carcinogenesis.

METHODSStreptavidin-peroxidase (SP) conjugated method was used to detect the expression of HCV NS(3) protein in NIH3T3 cells transfected with plasmid pRcHCNS(3)-5' and pRcHCNS(3)-3'. Telomerase activity was detected by an in situ telomerase activity labeling method, telomeric repeat amplification protocol polymerase chain reaction (TRAP-PCR) and telomerase PCR enzyme linked immunosorbent assay (ELISA) technology in the transfected and non-transfected NIH3T3 cells.

RESULTSHCV NS(3) protein was expressed in the NIH3T3 cells transfected with plasmid pRcHCNS(3)-5' expressing HCV NS(3) C-terminal deleted protein or with plasmid pRcHCNS(3)-3' expressing HCV NS(3) N-terminal deleted protein. The positive signal of HCV NS(3) protein was localized in the cytoplasm of NIH3T3 cells, and the signal intensity of the former was stronger. Telomerase activity in NIH3T3 cells transfected with plasmid pRcHCNS(3)-5' was stronger than that in NIH3T3 cells transfected with plasmid pRcHCNS(3)-3' (P < 0.01), whereas telomerase activity in NIH3T3 cells transfected with plasmid pRcCMV or untreated NIH3T3 cells was weaker than that in NIH3T3 cells transfected with plasmid pRcHCNS(3)-3' (P < 0.05). The expression level of HCV NS(3) protein was significantly correlated with the strength of telomerase activity (P < 0.05). The results obtained by in situ telomerase activity labeling corresponded to the results by telomerase PCR ELISA technology.

CONCLUSIONSHCV NS(3) protein may activate telomerase through endogenous mechanism to induce host cell transformation. The effect of HCV NS(3) C-terminal deleted protein on telomerase activity in the host cell may be stronger than that of HCV NS(3) N-terminal deleted protein. In situ telomerase activity labeling was a reliable technology for studying pathological morphology and telomerase activity in tissues and cells.

3T3 Cells ; Animals ; Enzyme-Linked Immunosorbent Assay ; methods ; Mice ; Plasmids ; genetics ; Polymerase Chain Reaction ; methods ; Telomerase ; genetics ; metabolism ; Transfection ; Viral Nonstructural Proteins ; genetics ; physiology

Objective To evaluate the additional role of 99mTc-MIBI SPECT-CT imaging before parathyroidectomy (PTX) for secondary hyperparathyroidism (SHPT) patients.Methods A total of 359 SHPT patients from the First Affiliated Hospital of Nanjing Medical University were enrolled in the study from April 2011 to January 2016.Patients were divided into two groups according to the difference of 99mTc-MIBI imaging techniques preoperatively.Patients in group A had dual-phase 99mTc-sestamibi parathyroid scintigraphy performed only.Patients in group B had SPECT-CT parathyroid scintigraphy added in the early phase.With the parathyroid glands confirmed by pathology after operation being the referencc,scnsitivity,specificity,consistency and surgical success rates of the patients in two groups were compared.Results 359 patients (166 men and 193 women) were enrolled,among which 339 on hemodialysis and 20 on peritoneal dialysis.The average age of those patients was (45.90± 11.20) years and median dialysis age was 84 (60.0,120.0) months.Total detection sensitivity,specificity,consistency in group A (169 patients) were 65.54％,56.52％,65.24％ and in group B (190 patients) were 73.84％,84.62％,74.21％ respectively.Total sensitivity and consistency were higher in group B than those in group A (P ＜ 0.01),while no difference in total specificity was observed between two groups (P=0.055).In group A,detection sensitivity of parathyroid glands were 47.56％,77.44％,57.14％ and 79.88％ in right upper gland (RU),right lower gland (RL),left upper gland (LU) and left lower gland (LL) respectively.Consistency in group A were 48.52％,77.51％,56.21％ and 78.70％ respectively.In group B,sensitivity were 58.33％,83.78％,69.78％ and 82.89％ in RU,RL,LU and LL respectively.Consistency in group B were 59.47％,84.21％,70.53％ and 82.63％ respectively.Sensitivity and consistency of upper glands were lower than lower glands in both groups (P ＜ 0.01).When SPECT-CT was added,sensitivity and consistency of upper glands for both sides were higher in group B than those in group A,while no difference of surgical success rates was found between two groups (87.57％ vs 92.63％,P=0.107).Conclusion 99mTc-MIBI SPECT-CT can be combined with anatomic image effectively and increase the sensitivity and consistency of total and upper parathyroid glands.It can also make the accurate location of the lesions,which improves the efficiency of the operation.

Objective To observe the characteristics of brain activation of multiple sclerosis (MS) patients and healthy controls at functional magnetic resonance imaging (fMRI) with olfactory stimulation,determine the locations of activation in areas of olfactory center and explore the MS olfactory related network.Methods Eighteen MS patients from the Affiliated Hospital of North Sichuan Medical College from February 2017 to March 2018 were enrolled as MS group,and 20 matched healthy adults during the same period served as controls.The Visual Analogue Scale was used to evaluate olfactory function in all subjects,the rest structure MRI was performed first,and volatile gases of lavender and rose solution were used to alternately stimulate olfactory during fMRI scanning.The brain activation was obtained by using matlab2013a and SPM8 softwares.The distribution and quantity of demyelination lesions were counted on T2 weighted image,and Spearman correlation analysis was done with SPSS 17.0 software package.Results The activated brain areas in the healthy control group included bilateral middle frontal gyrus,bilateral insula,bilateral supramarginal gyrus,bilateral orbitofrontal gyrus,right thalamus,right central anterior gyrus,bilateral cingulated gyrus,bilateral hippocampus,bilateral amygdala and bilateral superior frontal gyrus (t =2.11,P＜0.05).The activated brain areas in the MS group included right cerebellum,left insula,left superior temporal gyrus,right inferior frontal gyrus (t=2.19,P＜0.05).Compared with the control group,the MS group showed statistically significant decrease in activated values in right insula,right amygdala,right inferior frontal gyrus,right frontal middle gyrus,and the left supramarginal gyrus (t=2.04,P＜0.05).The distribution and number of demyelination lesions and major activation of brain regions with olfactory in the MS group demonstrated no significant correlation (r=-0.524,P=0.054).Conclusions Multiple brain areas involved in the olfactory processing and olfactory-related brain network existed.The activation of olfactory center had dominance in the right brain.The activation of the brain area in the MS group was significantly reduced,and the activation voxel and activation intensity were weakened.The olfactory-related brain network changed in MS patients.The distribution and number of demyelination lesions had no significant effect on the major activation of brain regions with olfactory stimulation.

Objective:To observe heart rate circadian rhythm in patients with chronic kidney disease (CKD) stage 5 and to analyze the effects of parathyroidectomy (PTX) on heart rate circadian rhythm in severe secondary hyperparathyroidism (SHPT) patients.Methods:A cross-sectional observation was performed in 213 patients with CKD stage 5 and 96 controls, and the patients were divided into those with severe SHPT (PTX group, n=70) and without severe SHPT (non-PTX group, n=143). Forty-six PTX patients were followed up prospectively. The baseline data were compared among these groups. Holter electrocardiogram was performed for each participant. Non-dipping heart rate was defined as night/day heart rate ratio greater than 0.9. Multiple linear regression analysis was used to analyze the related factors of heart rate circadian rhythm in patients with CKD stage 5. Results:The 24-hour, daytime and nighttime mean heart rate in patients with CKD stage 5 were all higher than those in controls, especially in PTX group (all P<0.05). The night/day heart rate ratios of controls and CKD stage 5 patients were (0.81±0.08) and (0.91±0.08) respectively ( P<0.01). Correlation analysis showed 24-hour and daytime or nighttime mean heart rate in patients with CKD stage 5 were positively correlated with serum levels of phosphorus and ln(alkaline phosphatase), while nighttime mean heart rate and night/day heart rate ratio were positively related with serum intact parathyroid hormone level. After adjusting with postoperative follow-up period (median time: 10.9 months), 24-hour and nighttime mean heart rate, and night/day heart rate ratio in PTX patients all decreased significantly (all P<0.01). Conclusions:Heart rate is increased and circadian rhythm is abnormal in patients with CKD stage 5, which are related with mineral and bone disorder. PTX significantly decreases 24-hour and nighttime mean heart rate in severe SHPT patients, and improves the heart rate circadian rhythm.