Objective　To investigate the expression of glial fibrillary acidic protein (GFAP) gene and its significance in the process of glioma cell differentiation induced by nordihydroguaiaretic acid (NDGA). Methods　Immunohistochemistry (IHC) and in situ hybridization were used to detect the expression changes of GFAP protein and GFAP mRNA qualitatively and quantitatively. Results　The expression levels of GFAP protein and GFAP mRNA in NDGA treatment group were significantly increased compared with the control group (P<0.01). Conclusion　NDGA could induce GFAP gene in malignant glioma cells and the up-regulation of this gene expression might be one of the mechanisms by which NDGA induces glioma differentiation.

Objective　To investigate the genomic methylation pattern of a malignant glioma cell line in the process of differentiation induced by nordihydroguaiaretic acid (NDGA). Methods　Methylation-sensitive arbitrarily primed PCR was used to study the genomic methylation changes. Results　Fragments of genomic DNA of PCR products in control groups digested with MspⅠ were smaller than those with HpaⅡ. No large fragment could be identified and at least three fragments of different sizes were demonstrated in the control group. In NDGA treatment group, comparing MspⅠ digestion with HpaⅡ digestion, the amount of PCR products was smaller with more DNA bands. The amount of PCR products in NDGA treatment group was increased with more DNA bands compared with that in the control groups. Conclusion　The genomic methylation level in SHG-44 cells was increased by NDGA in the differentiation process of SHG-44 cells. It suggests that genomic methylation pattern may be one of the targets for glioma cell differentiation induced by NDGA.

Objective To explore the relationship between chemokine receptor CXCR4 expression and invasiveness of pituitary adenomas. Methods A total of 78 pituitary adenoma specimens with comprehensive clinical data were surgically removed in Southwest hospital and 5 normal pituitary tissue samples were used as control. All the samples were fixed with 10 % formalin and then embedded with paraffin. Every case history was reviewed in detail. All the specimens were divided into invasive group (35 cases) and non-invasive group (43 cases) by the combined application of Knosp imaging diagnosis standard and Renzuyuan’s standard and referring to all operation records. The expressions of CXCR4 protein in 83 specimens were examined by immunohistochemistry. The relation between the expression of CXCR4 protein and invasiveness of the pituitary adenomas was analyzed. Results Immunohistochemistry showed that there was significantly high CXCR4 protein expression in invasive group and low in non-invasive group and normal pituitary tissue. There was a significant correlation between the expression level of CXCR4 protein and invasiveness of the pituitary adenomas (P005). Conclusion High CXCR4 protein expression is associated with the invasiveness of pituitary adenomas, suggesting that the expression of CXCR4 may have a value to assess the invasive pituitary adenomas.

OBJECTIVE To analyze for mutations of the PDS gene in patients with sensorineural hearing loss associated with enlarged vestibular aqueduct and analyze the molecular pathogenesis of enlarged vestibular aqueducts. METHODS Eighteen sporadic cases of large vestibular aqueduct syndrome and twelve control individuals with normal hearing were included in this study. Exons 6 and 9 of the PDS gene in all subjects were amplified by polymerase chain reaction and analyzed by direct DNA sequencing. RESULTS Analysis revealed 2 single base changes in exon 6 of one patient with large vestibular aqueduct syndrome. One was a G→C transversion at nucleotide position 611, and the other was a T→G transversion at nucleotide position 612, resulting in a predicted Gly→Ala substitution at position 204. No mutation in exons 6 and 9 of the PDS gene was found in the PDS gene of the control individuals. CONCLUSION Mutations of the PDS gene are responsible for the large vestibular aqueduct syndrome. Analysis of the PDS leftover sequence in patients with large vestibular aqueduct syndrome is the next step in elucidating the complicated causes of this disease.

Objective To establish rat model of impact spinal cord injury, observe the pathological changes of the model and assess its stability, reproducibility and consistency. Methods Moderate and severe spinal cord injury (SCI) models were established by using modified weight drop device. The pathological and functional changes after SCI were observed by means of BBB scoring, electrophysiology,immunohistochemistry and electronic microscopy so as to estimate the reproducibility of rat models and their consistency with severity of SCI. Results Locomotion and nerve impulse transduction along the spinal cord measured by motorial and sensory evoked potentials recovered gradually over time after SCI.However, the recovery rate of moderate SCI group was better than that of severe SCI group. Histological and immunohistochemical experiments showed that the glial scar as well as cavity were formed after SCI.Whereas, compared with moderate SCI group, the injury of severe SCI group was severer, with less spared tissue. Electronic microscopic observation displayed that hemorrhage, edema, neutrophilic granulocytic infiltration and chromatin margination of glia arose at day 1 after SCI. Vacuolization of mitochondria, degeneration of axon with edema could be seen at 2 weeks after injury. Degeneration of myelin and deposition of collagen fibril emerged at 8 weeks postinjury. Conclusions The rat models of impact SCI established in this study can distinguish the graded injury, and significantly correlate with the behavioral,electrophysiological and pathological outcomes, which indicates that the models possess good stability, reproducibility and consistency. Glial scar with cavity marked by GFAP or Vimentin is the pathological hallmark after SCI, and thereby GFAP or Vimentin can be used as a marker for demarcate the border of glial scar.

Objective To probe the diagnostic efficacy of fetal eehocardiography for characterizing complex congenital heart diseases.Methods Fetal echocardiography was performed on 49 cases of fetal complex congenital heart disease, the ultrasonic diagnosis was compared retrospectively with pathological results after autopsy.Results Antenatal sonographic diagnosis was in agreement with the pathological results in 42 cases (85.71 %), 7 cases were disagreed with pathological diagnosis (antenatal sonographic diagnosis was discrepancy in 3 cases, 4 cases were partially mis-classified).Twenty-four cases were combined with extra-cardiac malformations.Nine cases had chromosomal abnormality.Conclusions Fetal echocardiography is highly accurate for antenatal diagnosis of complex congenital heart disease, but it is hard to detect some type of cardiac anomalies.

Objective:To study the influence of different culture conditions on charcic and inhibition activity of nature killer cells ( NK) ,whether to join the modified K562 cells with IL-6 cytokine.Methods:According to the 5′end of the human IL-6 cDNA sequence,PCR primers designed to amplificate,express and transfect K562 cells cDNA library as a template for DNA.Genetic modified K562 cells as stimulating cells were prepared by expressing IL-6.To extract peripheral blood mononuclear cells( PBMC) from human peripheral blood.PBMC were explanted by genetic modified K562 stimulated.The expansion was initiated by CO-culture of PBMC and irradiate genetic modified K562 cell.The number of NK cell increased by directed induced generation of genetic modified K562 cell.Immunophenotypic analysis of NK cell surface markers was performed by flow cytometry (FCM).51Cr release assay was employed to measure the specific lysis skilling of NK cell target K562 cells.Results:We have constracted genetic modified K562 cells by genetic engineering.As stimulated cell added into the PBMC,an average of 760 ±18 fold expansion of CD56+CD16+CD3-cells was observed after 3 weeks of co-culture system.The NK cells population could proliferated more 91%±2% after expansion comparing with 6%± 0.4%in PBMC before expansion by FCM.The cytotoxical activity of NK cells which was induced by genetic modified K562 cell was the strongest than induced by IL-6 cytokine alone.The expanded NK cells lysed 92%±2% of K562 targets in a 5∶1 effector to target ratio.In this case,the NK cells induced by genetic modified K562 cells against tumor cells was more lethal.Conclusion:PBMC based in vitro expansion of natural killer cells was set up by genetic modified K562 cells.The cytotoxicity of NK cells was the strongest induced by genetic modified K562 cell treated.These results had important guiding significance for the the NK large number of amplification and used in clinical.

OBJECTIVE: To explore the expression of MEF2D in NPC tissues, study the relationship between the expression and prognostic. METHOD: Specimens from 101 NPC patients who were follow-up visited 1 to 7 years were analyzed for MEF2D by using immunohistochemistry. RESULT: (1) The expression of MEF2D was higher in the higher clinical stage. (2) Density and Grey of MEF2D was negative correlated (|r| = 0.865, P < 0.01). (3) NPC patients' survival rate after therapies was 52.5%, the survival curve of 1th clinical stage was higher than 4th. (4) The survival curves of MEF2D stages were no statistical significance. CONCLUSION There's statistical significance of the MEF2D expression in clinical stages, but not in survival curve, which indicated that MEF2D concerned with invasion and metastatic of NPC.
Adult ; Carcinoma ; Carcinoma, Squamous Cell ; metabolism ; pathology ; Female ; Humans ; Lymphatic Metastasis ; MADS Domain Proteins ; metabolism ; MEF2 Transcription Factors ; Male ; Middle Aged ; Myogenic Regulatory Factors ; metabolism ; Nasopharyngeal Carcinoma ; Nasopharyngeal Neoplasms ; metabolism ; pathology ; Neoplasm Staging ; Prognosis

Objective:Through the establishment and application of the biosafety autopsy pathology platform in Huoshenshan Hospital, the feasibility and application effect of the biosafety autopsy pathology platform were analyzed.Methods:The feasibility and application effect of the biosafety autopsy pathology platform were analyzed by layout design, instruments and equipment preparation, testing methods examination, and effect evaluation.Results:A total number of 26 cases of systematic autopsy and 8 cases of minimally invasive autopsy (puncture) were performed on the biosafety autopsy pathology platform, and no one was infected. Some pathology original findings were identified, including COVID-19 and pathological characteristics of identification, SARS respiratory failure mechanism and treatment significance, systemic distribution and spreading mechanism of the new coronavirus, the " storm" of inflammation pathological basis, some tumor markers rise in pulmonary pathological cell source and the overcast with fibrosis characteristics, such findings play important roles in the clinical diagnosis and treatment of COVID-19.Conclusions:The study of autopsy pathology is of great significance for the prevention and control of emerging infectious disease, which calls for early intervention. To promote the standard construction of biosafety autopsy platform is the key to the pathological study of emerging infectious diseases. Pathological research and clinical diagnosis and treatment should be combined to inform each other.

Objective To investigate the clinical feasibility and effectiveness of 3-D printing (3DP) combined with 3-D laparoscopic nephron-sparing surgery (LNSS) for partial endogenous renal cell carcinoma.Methods A retrospective analysis was made of the clinical data of 79 patients with partial endogenous renal cell carcinoma who were admitted to our department from July 2015 to October 2018.There were 46 males and 33 females.Their average age was (50.9 ± 7.9) years old,ranged from 33 to 68 years old.Tumor stages were T1aN0M0 in 53 cases and T1bN0M0 in 26 cases.The preoperative serum creatinine ranged from 40 to 107 μmol/L,with an average of (72.4 ± 14.2) μmol/L.The preoperative GFR ranged from 19 to 54 ml/min,with an average of (40.2 ± 6.2) ml/min.Thirty-four patients underwent 2-D laparoscopic nephron-sparing surgery (2DLNSS) based on preoperative enhanced CT scans.Forty-five patients underwent 3-D printing (3DP) based on three-dimensional reconstruction of renal CT scans.Seventeen patients underwent 2-D laparoscopic nephron-sparing surgery guided by 3-D printing model(3DP-2DLNSS),and 28 patients underwent 3-D laparoscopic nephron-sparing surgery guided by 3-D printing (3DP-3DLNSS).Serum creatinine levels ranged from 42 to 122 μmol/L with an average of (86.3 ± 14.8) μmol/L,and creatinine levels ranged from 8 to 66 μmol/L with an average of (19.1 ± 14.1) μmol/L.Six months after operation,the GFR of the kidney was 9-36 ml/min with an average of (21.4 ± 6.4)ml/min,and the fluctuation range was 6-40 ml/min with an average of (19.2 ± 8.8) ml/min.There was no statistical difference in the incidence of complications and pathological types after operation.Results There was no statistical difference in general data of preoperative patients.In intraoperative and post-operative statistics,the time of exploring renal artery was shorter than that of 2DLNSS (33.7 ± 7.5) min in 3DP-2DLNSS (28.3 ± 8.2,P =0.015) min and 3DP-3DLNSS (27.8 ± 6.5,P =0.002) min.In tumor detection time,3 DP-2DLNSS was shorter than 2DLNSS group (41.2 ± 6.6 vs.46.5 ± 6.9 min,P =0.012),and 3 DP-3DLNSS was shorter than 3DP-2DLNSS (35.4 ± 7.3 vs.41.2 ± 6.6 min,P =0.009).In warm ischemia time,3DP-2DLNSS min was shorter than 2DLNSS (23.5 ±9.7 vs.33.9 ±7.5 min P ＜0.001),and 3DP-3DLNSS was shorter than 3DP-2DLNSS (18.3 ± 4.6 vs.23.5 ± 9.7,P =0.023).In surgical time,3DP-2DLNSS (115.7 ± 23.0) min and 3DP-3DLNSS (103.3 ± 22.8) min were shorter than 2DLNSS (132.4 ± 28.9) min (P =0.031,P ＜ 0.001).In intraoperative bleeding volume,3 DP-3 DLNSS was less than 2DLNSS (117.9 ± 17.9 vs.130.6 ± 16.8,P =0.009) ml.Fasting for 1 to 4 days after operation,with an average of (1.7 ± 0.8) days.The indwelling catheterization ranged from 1 to 8 days after operation,with an average of (3.9 ± 1.3) days.Negative pressure drainage was removed 2-9 days after operation,with an average of (4.9 ± 1.4) days.And the hospitalization 5-11 days after operation,with an average of (7.3 ± 1.5) days.Conclusions Preoperative 3D printing combined with intraoperative 3D laparoscopic nephron sparing surgery for partial endogenous renal tumors is safe and effective,which is superior to the previous CT scan alone and intraoperative 2D laparoscopic treatment.