[Analysis of the difference in MGMT promoter status in gliomas and its significance in prognosis assessment].

The data of 1 268 newly diagnosed gliomas from the Fourth Ward of Neurosurgery Department of Beijing Tiantan Hospital between April 2013 and March 2022 were retrospectively analyzed. Based on postoperative pathology, the gliomas were divided into groups: oligodendrogliomas (n=308), astrocytomas (n=337) and glioblastomas (n=623). According to the O6-methylguanine-DNA methyl transferase (MGMT) promoter status defined by the 12% of best cut-off value in previous research results, patients were divided into methylation group (n=763) and non-methylation group (n=505). Methylation level [M (Q1, Q3)] in patients with glioblastoma, astrocytoma and oligodendroglioma was 6% (2%, 24%), 17% (10%, 28%) and 29% (19%, 40%), respectively (P<0.001). Compared with non-methylation patients, the progression-free survival (PFS) and overall survival (OS) of glioblastomas patients with methylation of MGMT promoter demonstrated more favorable prognosis [M (Q1, Q3)]) of PFS: 14.0 (6.0, 36.0) months vs 8.0 (4.0, 15.0) months, P<0.001; M (Q1, Q3) of OS: 29.0 (17.0, 60.5) months vs 16.0 (11.0, 26.5) months, P<0.001]. In astrocytomas patients, the PFS was much longer for those with methylation [the median PFS of patients with methylation was not observed at the end of follow-up, but those without methylation showed a median PFS of 46.0 (29.0, 52.0) months] (P=0.001). However, no statistically significant difference was observed in OS [the median OS of patients with methylation was not observed at the end of follow-up, but those without methylation had a median OS of 62.0 (46.0, 98.0) months] (P=0.085). In oligodendrogliomas patients, no statistically significant differences of PFS and OS were observed between patients with methylation and those without methylation. MGMT promoter status was a related factor affecting PFS and OS in glioblastomas (PFS: HR=0.534,95%CI: 0.426-0.668, P<0.001; OS: HR=0.451, 95%CI: 0.353-0.576, P<0.001). Moreover, MGMT promoter status was also a related factor affecting PFS in astrocytomas (HR=0.462, 95%CI: 0.221-0.966, P=0.040), but not for OS (HR=0.664, 95%CI: 0.259-1.690, P=0.389). The methylation level of MGMT promoter differed substantially in different types of gliomas, and the status of MGMT promoter profoundly affected the prognosis of glioblastomas.

[Clinical characteristics of 13 cases with entrapped temporal horn syndrome and efficacy of refined temporal-to-frontal horn shunt].

Thirteen consecutive patients with entrapped temporal horn syndrome in the Department of Neurosurgery of Beijing Tiantan Hospital from February 2018 to September 2022 were retrospectively analyzed, and there were 5 males and 8 females, with a mean age of (43±21) years. Increased intracranial pressure caused by hydrocephalus was the main clinical symptom. All the patients underwent refined temporal-to-frontal horn shunt, and all the symptoms were improved after surgery. Postoperative Karnofsky performance score (KPS) [90 (90, 100)] was higher than preoperative KPS [57 (40, 70)] (P=0.001). However, postoperative entrapped temporal horn volume [13.85 (8.90, 15.25) cm3] decreased, compared with preoperative volume [66.52 (38.65, 88.65) cm3] (P=0.001). Likewise, postoperative midline shift [0.77 (0, 1.50) mm] was longer than preoperative midline shift [6.69 (2.50, 10.00) mm] (P=0.002). No surgery-related complications were observed after the operation. Therefore, the refined temporal-to-frontal horn shunt is safe and effective treatment for entrapped temporal horn syndrome, with favorable outcomes.

[The outcomes of Tiantan first-aid protocol on critically ill patients with primary central nervous system lymphoma].

Objective: To examine the outcomes of Tiantan first-aid protocol on critically ill patients with primary central nervous system lymphoma (PCNSL). Methods: The clinical data of 18 patients with PCNSL who were treated according to Tiantan first-aid protocol at Department of Neurosurgery,Beijing Tiantan Hospital, Capital Medical University from November 2019 to December 2021 were retrospectively analyzed. There were 9 males and 9 females, aged (56.9±11.1)years (range: 29 to 77 years). The median Karnofsky performance status(KPS) score at admission was 40 (range: 20 to 60). Three patients were mild coma, 3 were lethargy and 12 were conscious. The mean midline shift was 0.7 cm (range: 0 to 1.8 cm). After admission, all patients were treated according to the plan of rapid biopsy, rapid routine pathology and rapid salvage chemotherapy. The treatment procedures, clinical and radiographic outcomes, KPS score and adverse reactions of patients after chemotherapy were collected. Results: All of the 18 patients completed the first-aid treatment. The median duration from admission to the biopsy was 1 day (range: 0 to 5 days), from biopsy to routine pathological diagnosis was 1 day (range: 1 to 4 days) and from routine pathology to salvage chemotherapy was 1 day (range: 0 to 4 days). All the patients were pathologically confirmed with diffuse large B cell lymphoma, 1 patient was double-hit lymphoma. Seventeen patients underwent clinical remission and 1 died of cardiac dysfunction. The successful salvage rate was 17/18. Radiologically, complete remission was observed in 1 case, partial remission in 16 cases, and stable disease in 1 case. The median KPS score at discharge was 60 (range: 30 to 80). The mild gastrointestinal, hematological and hepatic adverse effects were observed after chemotherapy. Conclusion: Tiantan first-aid protocol is effective for critically ill patients with PCNSL, which has the merit to be popularly used and improved.

[Different treatment regimens for primary central nervous system lymphoma:based on SEER database].

Objectives: To explore the prognostic factors of primary central nervous system lymphoma(PCNSL) and to analyze the efficacy of different treatment methods. Methods: Clinical data of 4 812 patients with PCNSL in SEER database from January 1975 to December 2016 were retrospectively analyzed.Among them, 2 831 were male and 1 981 were female, the ratio of male to female was 1.4∶1.0.There were 2 236 cases(46.47%) under 60 years old, 1 718 cases(35.70%) aged 60 to 74 years old, and 858 cases(17.83%) aged 75 years old or above. Two thousand four hundred and seventeen cases(50.23%) had supratentorial tumors, 299 cases (6.21%) had infratentorial tumors, and 554 cases(11.51%) had multiple brain tumors, 1 542 cases (32.04%) were other or unspecified location.Three thousand five hundred and thirteen cases(73.00%) had diffuse large B-cell lymphoma (DLBCL), 234 cases(4.86%) had non DLBCL, 1 065 cases (22.13%) had other or unspecified types of tumor.The treatment included 2 011 cases (41.77%) of biopsy, 61 cases (1.27%) of subtotal resection(STR), 54 cases (1.12%) of gross total resection(GTR), 2 384 cases (49.54%) of biopsy and chemotherapy, 159 cases (3.30%) of STR and chemotherapy, 144 cases (3.00%) of GTR and chemotherapy.Univariate and multivariate Cox regression models were used to analyze the prognostic factors affecting the overall survival of the patients.Fine-Gray test and competitive risk model were used to analyze the prognostic factors affecting cancer-specific survival.Kaplan-Meier method and Log-rank test was used for survival analysis. Results: Univariate and multivariate Cox regression analyses showed that age, race, marital status, tumor site, pathological subtype, surgery, chemotherapy, combined with other malignant tumors, and HIV infection were the independent prognostic factors affecting the overall survival of PCNSL patients.The results of Fine-Gray test and competitive risk model analyses showed that age, race, marital status, tumor location, pathological subtype, surgical method, chemotherapy, combined with other malignant tumors, and HIV infection were independent prognostic factors affecting cancer-specific survival, while gender and radiotherapy had no significant correlation with cancer-specific survival.Compared with biopsy, PCNSL patients may benefit from surgical resection (STR:HR=0.805, 95%CI:0.656‒0.989, P=0.04; GTR:HR=0.521, 95%CI:0.414‒0.656, P<0.01).Kaplan-Meier survival analysis showed that the median survival time of biopsy+chemotherapy group was 28 months (95%CI:24.497‒31.503), 2 months (95%CI:1.756‒2.244) in the biopsy group, 2 months (95%CI:1.410-2.590) in the STR group, 19 months (95%CI:0‒39.311) in the biopsy+chemotherapy group, 67 months (95%CI:46.187-87.813) in the STR+chemotherapy group, 84 months (95%CI:57.448‒110.552) in the GTR+chemotherapy group.The median survival time of patients with different treatment methods was statistically significant (P<0.01). Conclusions: Surgical resection may improve the prognosis of some PCNSL patients.Patients who have access to receive GTR or STR combined with chemotherapy may have prolonged Cancer-specific survival.

[Induction of hepatocellular carcinoma in B6C3 (F1) mice chronicly exposed to trichloroethylene with enhanced acetylation of histone H2AK9ac and SET expression in the liver tissue].

Objective: To establish an animal model of trichloroethylene (TCE) -induced liver cancer following chronic exposure and to understand the changes in SET expression and histone acetylation, potentially serving as a molecular mechanism for TCE-induced hepatocarcinogenesis. Methods: B6C3 mice at 6 weeks were treated with TCE at a series of doses (500, 1000 and 2000 mg/kg) by gastric gavage, with corn oil used as the negative control and carbon tetrachloride (CCl(4)) as the positive control. The serum and liver were sampled for the determination of biochemical indexes and pathological examination after 56 weeks of chemical exposure. Western blot was used to determine the levels of SET, H2AK9ac and HDAC1 expression. Results: The overall survival rate of the mice in various groups was 90.4% (141/156) , with no statistical difference between groups (P>0.05) . Compared with the negative control, the organ coefficient for the liver in the high dose TCE group and the positive control group were significantly increased (P<0.05) . The levels of ALT, AST, LDH and BUN in the all the three TCE groups and the positive control were significantly higher than those in the negative control (P<0.01) . CREA levels in the 1000 and 2000 mg/kg TCE groups were significantly higher than those in the negative control (P<0.05) . Statistical increases in the incidence of hepatocellular carcinoma and the activities of ALT and AST in various doses of TCE-exposed mice as compared with the control were observed (P<0.01) , in a dose-dependent manner. In the 1000 and 2000 mg/kg of TCE treated mice, levels of SET and H2AK9ac were increased (P<0.05) , while HDAC1 was decreased (P<0.05) , Compared to the tissue adjacent to liver cancer, in the 1000 and 2000 mg/kg TCE groups, the levels of SET were increased (P<0.05) , while HDAC1 was decreased (P<0.05) , and H2AK9ac increased in the 2000 mg/kg group. Conclusion: The hepatocellular carcinoma mouse model induced by chronic exposure to trichloroethylene was successfully established, with enhanced SET protein expression and H2AK9ac in the hepatic tissue.

MicroRNA-206 inhibits tumor metastasis of nasopharyngeal carcinoma through targeting G6PD.

Nasopharyngeal carcinoma (NPC) is a main type of otolaryngological malignancy. In many cancers, miR-206 functions as a tumor suppressor, suppressing cell proliferation, migration and invasion. The purpose of this study was to explore how miR-206 worked on cell metastasis in NPC. The mRNA levels of miR-206 and G6PD were determined in NPC tissues and cell lines by RT-qPCR and Western blot. Transwell assay was applied to evaluate the migratory and invasive capacities. Dual luciferase reporter assay was employed to confirm that miR-206 mediated the expression of G6PD in C666-1 cells. In this study, miR- 206 was downregulated, whereas G6PD was upregulated in NPC tissues and cell lines. In addition, G6PD was identified as a direct target gene of miR-206 in C666-1 cells. The expression of G6PD was mediated by miR-206, which could partially reverse the inhibitory effects of miR-206 on the migration, invasion and EMT in C666-1 cells. In conclusion, miR-206 regulated the migratory, invasive and EMT abilities through directly targeting the 3'-UTR of G6PD mRNA in C666-1 cells. The newly identified miR-206/G6PD axis provides novel insight into the pathogenesis of nasopharyngeal carcinoma.

[Factor analysis of cerebrospinal fluid spread in glioblastoma].

Objective: To analyze the prognosis factors of cerebrospinal fluid (CSF) spread after surgery in glioblastoma (GBM) patients when tumors progressed and the effect factors on prognosis. Methods: A retrospective study was conducted on 124 patients who were pathologically diagnosed as glioblastoma after surgery, and found tumor progressed during regularly follow-up at Department of Neurosurgery, Beijing Tiantan Hospital, Capital Medical University between January 2009 and August 2017.There were 82 males and 42 females, aged 47.9 years(range: 19 to 75 years) .Patients were divided into local recurrence group(96 cases) and CSF spread group (28 cases) .Clinical data were recorded in detail and compared by independent sample t test or χ(2) test.Kaplan-Meier survival curves was used to demonstrated the distribution of progression free survival (PFS) overall survival (OS) and post progression survival (PPS), and differences between local recurrence and CSF spread groups were assessed by Log-rank test.Cox proportion hazard regression analysis was used to identify independent prognostic factors. Results: Logistics regression analysis showed ventricle entry was the only prognosis factor of CSF spread (OR=2.667, 95% CI: 1.128 to 6.304, P=0.025).No significant distinction was observed in PFS between CSF spread group and local recurrence group(7.0 months vs.9.3 months, P=0.066).However, OS and PPS were substantially shortened in CSF spread group (13.0 months vs.23.0 months, P=0.011; 6.0 months vs.11.0 months, P=0.022, respectively).Mutations of isocitrate dehydrogenase gene, distant spread, gross-total resection, Ki-67 index>30% were independent prognostic factors of GBM patients. Conclusions: Ventricle entry is a prognosis factor for CSF spread, after which the median OS and PPS are markedly diminished.However, ventricle entry is not independent prognosis factor shortening survival.

[Identification of circulating tumor cells in peripheral blood for gliomas by detection of aneuploid cells].

Objective: To investigate the feasibility of detecting circulating tumor cells based on capture of heteroploid chromosome cells in peripheral blood of glioma patients. Methods: A total of 88 patients who were considered to suffer from gliomas and 10 healthy volunteers were enrolled in this study during January 2016 to December 2016 at Beijing Tiantan Hospital, from whom 6 ml preoperative blood was collected. Subtraction enrichment (SE)-immunostaining FISH (iFISH) was applied to capture the heteroploid chromosome 8 cells in those samples. Meanwhile, centromere probe 8(CEP-8)-FISH was used to identify aneuploid cells in 10 tumors and 10 brain tissues. Results: Numerous heteroploid chromosome 8 cells were observed in tumors whereas negative result was present in brain tissues (P<0.01). CTC was successfully detected in 90.9% glioma patients, in contrast, only one healthy volunteer was shown with a heteroploid chromosome 8 cell (P<0.01). Glial fibrillary acidic protein was not exhibited in the overwhelming majority of CTC (96.1%). High grade glioma (HGG) without IDH mutation possessed much more CTC than low grade (12.0 vs 2.2), P<0.01. Furthermore, multiploidy (≥5 copies) CTC accounted for a much significant percentage in HGG, either in tumors originating from oligodendrocyte or astrocyte (75.9% vs 56.0%), P<0.01; 62.7% vs 51.7%, P=0.016, respectively). Conclusion: CTC could be identified and enumerated in glioma by detecting aneuploidy cells in blood. The number and multiploidy proportion of CTC may be correlative with tumor grade and molecular characteristics.

[Association between dietary magnesium and the risk of type 2 diabetes in Harbin residents].

Objective: To investigate the relationship between the magnesium intake and patterns of diary and the risk of type 2 diabetes in Harbin residents. Methods: On April 2010, 24 communities in 7 districts of Harbin were selected as research sites using multi-stage stratified random cluster sampling method. A total of 9 734 residents aged 20-74 years was investigated using general questionnaire survey, dietary survey and biochemical indicators test. A total of 9 376 subjects were included in the study. Factor analysis was used to analyze dietary patterns. According to the quartile of dietary magnesium intake, the subjects were divided into four groups, from Q(1) to Q(4) group. The multivariate logistic regression model was used to analyze the relationship between dietary magnesium intake and the risk of type 2 diabetes within different dietary patterns. Results: A total of 998 subjects with type 2 diabetes were screened. The median age of the diabetic group and the non-diabetic group were 54.8 and 50.8, respectively, and the males accounted for 43.4% (2 896 cases) and 34.6% (433 cases), respectively. The magnesium intake median (P(25), P(75)) of two groups was 336.36 (257.31, 440.65) and 339.50 (264.51, 443.78) mg/d. Four dietary patterns were identified as western dietary mode, savvy dietary mode, traditional dietary mode, and staple food mode. In the western dietary model, the Q(4) group had a higher risk of type 2 diabetes than Q(1) group, with an OR (95%CI) value of 1.56 (1.06 to 2.32). However, in the savvy diet mode, compared with the Q(1) group, the risk of diabetes in the Q(4) group was lower, and the OR (95%CI) value was 0.61 (0.37 to 0.96). There was no statistically significant association between dietary magnesium intake and the risk of type 2 diabetes without considering dietary patterns (P>0.05). Conclusion: Dietary magnesium intake has a different relationship with the risk of type 2 diabetes within different dietary patterns.

[Clinical prognostic factors of adult supratentorial glioblastoma].

Objective: To analyze the treatment effect of patients with glioblastoma (GBM) and explore prognostic factors. Methods: The clinical data of 635 patients diagnosed as GBM at Neurosurgical Oncology Department Ⅳ of Beijing Tiantan Hospital, Capital Medical University from January 2007 to March 2018 were retrospectively reviewed. There were 386 males and 249 females with an age of (48.7±11.8) years (range: 18-75 years). Patients were divided into three groups according to the time of admission: 2007-2010 group(n=174), 2011-2014 group (n=237) and 2015-2018 group (n=224). Kaplan-Meier plot was used to analyze the effects of different treatment periods, treatment schemes and clinical factors on the survival of patients with GBM. Cox proportion hazard regression analysis was used to identify independent prognostic factors. Results: The median progression-free survival (PFS) and overall survival (OS) of patients in 2007-2010 group, 2011-2014 group, 2015-2018 group was 9.0 months (95% CI: 7.5-10.5), 10.0 months (95% CI: 8.8-11.2), 12.0 months (95% CI: 10.7-13.3) and 17.0 months (95% CI: 13.2-20.8), 20.0 months (95% CI: 16.9-23.1), 23.0 months(95% CI: 17.5-28.5), respectively. The PFS and OS of patients improved significantly over the years (χ(2)=9.693, P=0.008 and χ(2)=8.616, P=0.013). Multivariate survival analysis showed that age, extent of resection, radiotherapy and tumor distant dissemination were independent prognostic factors (all P<0.05). Conclusions: With the continuous development of clinical treatment regimen, the therapeutic effect of Chinese GBM patients has improved remarkably. Age, extent of resection, radiotherapy and tumor distant dissemination are independent prognostic factors associated with survival time.

[Analysis of the potential role of SET in chromium-induced malignant transformation cells based on quantitative proteomics].

Objective: To investigate alteration of proteins profile in malignant transformation bronchial epithelial cells(16HBE-T) induced by hexavalent chromium[(Cr(VI))] and analyze the expression level of SET protein, then to provide some new insights for the carcinogenesis mechanism of Cr(VI). Methods: Total protein was extracted from 16HBE cells and was alkylated and desalinated before digested into peptides. The products were labeled with Tandem Mass Tag (TMT) and identified using LC-ESI-MS/MS. Results: A total of 3 517 proteins were found, expression differences greater than 1.5 or less 0.67 times were to found have 185 and 201 proteins, respectively. Gene enrichment analysis revealed that differential proteins were mainly involved in autophagy, DNA damage repair, RNA processing and other biological processes. Western blot results showed the expression level of SET was significantly increased while downregulated in histone H3K18/27 acetylation and p53 protein. Conclusion: Proteins involved in multiple biological processes altered in 16HBE-T cells and regulation mode of SET inhibiting histone H3K18/27 acetylation regulating transcriptional activity of p53 may paly an important role in Cr(VI)-association carcinogenesis.

[The application of cortical and subcortical stimulation threshold in identifying the motor pathway and guiding the resection of gliomas in the functional areas].

Objective: This study aimed to analyze the application of cortical and subcortical stimulation threshold in identifying the motor pathway and guiding the resection of gliomas in the functional area, and to illustrate the minimal safe threshold by ROC method. Methods: Fifty-seven patients with gliomas in the functional areas were enrolled in the study at Beijing Tiantan Hospital from 2015 to 2017. Anesthesia was maintained intravenously with propofol 10% and remifentanil. Throughout the resection process, cortical or subcortical stimulation threshold was determined along tumor border using monopolar or bipolar electrodes. The motor pathway was identified and protected from resection according to the stimulation threshold and transcranial MEPs. Minimal threshold in each case was recorded. Results: Total resection was achieved in 32 cases(56.1%), sub-total resection in 22 cases(38.6%), and partial resection in 3 cases(5.3%). Pre-operative motor disability was found in 9 cases. Compared with pre-operative motor scores, 19 exhibited impaired motor functions on day 1 after surgery, 5 had quick recovery by day 7 after surgery, and 7 had late recovery by 3 months after surgery. At 3 months, 7 still had impaired motor function. The frequency of intraoperative seizure was 1.8%(1/57). No other side effect was found during electronic monitoring in the operation. The ROC curve revealed that the minimal safe monopolar subcortical threshold was 5.70 mA for strength deterioration on day 1 and day 7 after surgery. Univariate analysis revealed that decreased transcranial MEPs and minimal subcortical threshold ≤5.7 mA were correlated with postoperative strength deterioration. Conclusions: Cortical and subcortical stimulation threshold has its merit in identifying the motor pathway and guiding the resection for tumors within the functional areas. 5.7 mA can be used as the minimal safe threshold to protect the motor pathway from injury.

[Studies on the DNA damage in the transformed bronchial epithelial cells induced by hexavalent chromium].

Objective: To investigate DNA damage in the transformed human bronchial epithelial cells (16HBE) induced by hexavalent chromium (Cr(6+)) and further elucidate the potential carcinogenesis mechanism of Cr(6+). Methods: 16HBE were treated with different concentration of Cr(6+ ()0, 0.625, 1.25, 2.5 µmol/L) for 15 weeks. The malignant degrees of transformed cells were identified by the assays for anchorage-independent growth and tumorigenicity. According to the single cell gel electrophoresis (SCGE) assay, the DNA damage rate was calculated. The expression level of 53BP1 was determined by Western blot. Results: Chromium-treated cells could form colonies in soft agar and tumors in nude mice. Compared with the control group, colony formation efficiency of 1.25µmol/L and 2.5 µmol/L Cr(6+)-treated cells in soft agar showed significant increases (p<0.05) . The 2.5 µmol/L Cr(6+)-treated cells also formed tumors subcutaneously in nude mice. Cr(6+) could cause different degree of DNA damage to 16HBE cells in a dose-dependent manner. In addition, Western blot analyses showed that 53BP1 was aberrantly down-regulated at 2.5 µmol/L dose and has no significant changes at 0.625 µmol/L and 1.25 µmol/L dose under the treatment of Cr(6+). Conclusion: The declined expression of 53BP1 may mediate Cr(6+)-induced DNA damage and further involved in the cell malignant transformation.

[Trichloroethylene-induced abnormal methylation on promoter region of SET in hepatic L-02 cells].

Objective: To explore the trichloroethylene-induced alteration of methylation on the promoter region of SET and related mechanisms in hepatic L-02 cells. Methods: L-02 cells were treated with different concentrations of TCE(0 mmol/L, 1 mmol/L, 2 mmol/L, 4 mmol/L, 8 mmol/L) for 24 h. The genomic DNA were then extracted and modified by bisulfite sodium. The DNA methylation was then analyzed using bisulfite sequencing PCR (BSP). Results: The overall methylation on promoter region of SET was decreased along with the increased concentrations of TCE in hepatic L-02 cells. Moreover, 73 CpG islands were found abnormally altered, among which 9 were predicted in transcriptional factor binding regions. Conclusion: The decreased levels of CpG islands in the transcriptional factor binding region may contribute to the elevation of SET in TCE-induced hepatotoxicity.

Relationship between transmembrane serine protease expression and prognosis of esophageal squamous cell carcinoma.

Esophageal squamous cell carcinoma is the most common type of esophageal cancer in Eastern Europe and Asia, being the 6th most common cause of cancer deaths worldwide. The aim of this study was to analyze the expression of transmembrane serine protein in esophageal squamous cell carcinoma, and to correlate it with the clinical biological features of esophageal cancer. The expression of transmembrane protease serine 4 (TMPRSS4) mRNA and protein in carcinoma tissues and corresponding adjacent tissues and non-tumorous esophageal tissues was determined using PCR (qRT-PCR). The results show that both TMPRSS4 mRNA and protein expression were remarkably lower in adjacent normal tissues than in tumorous tissues. TMPRSS4 protein expression in esophageal carcinoma was correlated with patient demographic characteristics, tumor type, high TNM stages and overall survival (OS). Based on the experimental results, we conclude that TMPRSS4 is closely related to the occurrence, development and metastasis of esophageal squamous cell carcinoma.

[Foundation of preoperative prognosis estimation model for glioblastoma multiforme].

Objective: This study explored the preoperative prognostic factors of patients with glioblastoma multiforme (GBM) in order to propose a preoperative prognosis estimation model. Methods: The clinical data of 416 patients diagnosed with GBM in Beijing Tiantan Hospital affiliated to Capital Medical University from 2008 to 2015 were retrospectively reviewed.A total of nine factors: gender, age, duration of symptoms, preoperative epilepsy, preoperative muscle weakness, preoperative headache, preoperative KPS score, tumor location and tumor diameter were enrolled in the survival analysis.The significant factors identified by Kaplan-Meier plot were further collected in the multivariate Cox regression analysis.On the basis of multivariate analysis results, a preoperative prognosis estimation model was founded. Results: Univariate analysis showed that Age ≥50 years, without preoperative epilepsy, tumor located in non-frontotemporal lobe, tumor diameter ≥6 cm and preoperative KPS score <70 were prognostic risk factors (P<0.05). Multivariate analysis revealed that Age ≥50 years, without preoperative epilepsy, tumor located in non-frontotemporal lobe were independent risk factors (P<0.05). The prognostic estimation model based on the independent risk factors divided the whole cohort into three subgroups with different survival (P<0.001). Conclusions: The more risk factors, the higher score but poorer prognosis. Patients in the high-risk group had lower gross total resection degree but higher rate of postoperative complications, which suggested that aggressive resection was not suitable for high-risk patients.

[An investigation of trichloroethylene-induced effects on histone methylation in L-02 hepatic cells].

Objective: To further explore TCE-induced hepatotoxicity and its mechanisms by identification of trichloroethylene (TCE) induced abnormal histone methylation in human liver cells. Methods: L-02 cells were treated with 0 and 8 mmol/L TCE for 24 h. Histones were extracted by acid. Liquid chromatography electrospray ionization tandem mass spectrometry (ESI-LC-MS/MS) were used to identify and quantify TCE related histone methylations. TCE induced abnormal methylation of H3K79 me2 and H3K79 me3 were validated by Western blot analysis. The further analysis of the function of histone abnormal methylation modifications were done by single cell gel electrophoresis (SCGE) and Western blot analysis of p53 and ɤH2AX. Results: After treatment with TCE for 24 h in L-02 cells, the 36 TCE related histone methylation sites in 28 peptide segments were identified by MS. After treatment with TCE in concentrations of 0 and 8.0 mmol/L in L-02 cells for 24 h, the relative expression level of histone H3K79 me3 were 1.00±0.06, 0.70±0.09 (t=15.01, P=0.015); the relative expression level of histone H3K79 me2 were 1.00±0.05, 0.74±0.07 (t=16.69, P=0.018); the Olive Tail Moment about DNA damage were 1.46±0.28, 3.12± 0.68 (t=15.22, P=0.018); the relative expression levels of p53 were 1.00±0.04, 1.24±0.04 (t=18.71, P= 0.012); and the relative expression levels of ɤH2AX were 1.00 ± 0.03, 1.56 ± 0.11 (t=8.32, P=0 045). Conclusion: TCE can induce changes in the relative expression level of H3K79 me2 and H3K79 me3 in L-02 cell, and induce DNA damage, suggesting that TCE may induce changes in the relative expression level of H3K79 me2 and H3K79 me3 by DNA damage.

Spinal sample showing p-JNK and P38 associated with the pain signaling transduction of glial cell in neuropathic pain.

OBJECTIVE: To investigate the signaling pathways after astrocytes were activated in neuropathic pain. METHODS: Thirty-six Sprague Dawley (s.d.) rats were randomly divided into two groups (each group with 18 s.d. rats) including chronic constriction injury (CCI) of the sciatic nerve model group and sham operation group. Operation was perform ed on the right leg in all rats. The lumbar spin al cord (L4 and L5) was taken to make paraffin slices on the 1st day before operation and the 1st, 3rd, 7th, 14th and 28th day after operation in each group. Paraffin slices were labeled with p38 mitogen-activated protein kinase (p38MAPK) and c-Jun N-terminal kinase (JNK) by immunofluorescence staining, and then were co-labeled with hexaribonucleotide binding protein-3 (NeuN), glial fibrillary acidic protein (GFAP) and anti-integrin αM (CD11b) antibody (OX-42) to explore the associations of p38MAPK and JNK with nerve cells or glial cell. RESULTS: Compared with sham group, the pain threshold was significantly decreased, and astrocyte-activated markers, GFAP and vimentin were significantly increased in CCI group. The mean fluorescence intensities of p38MAPK and JNK were increased in the right spinal dorsal horn of CCI group. The coexpression of JNK and GFAP was found in astrocytes of the spinal dorsal horn in CCI group. CONCLUSION: JNK signal transduction pathway is involved in the pain signaling transduction of astrocytes.

Hypolipidemic effect of safflower yellow and primary mechanism analysis.

We examined the hypolipidemic effect of safflower yellow (SY) on hyperlipidemic mice and its influence on the biological synthesis of cholesterol in cells. Over 4 weeks, the levels of total cholesterol, triglyceride, low-density lipoprotein cholesterol, and high-density lipoprotein cholesterol in serum were detected using a kit; mouse liver samples were acquired for paraffin sections, and mouse liver cells were observed under light microscope. Chinese hamster ovary cells were cultured in vitro, and an amphotericin B-cell model was adopted to observe the inhibitory effect of SY on the biological synthesis of intracellular cholesterol. An enzyme-linked immunosorbent assay was used to detect the survival rate of Chinese hamster ovary cells. The middle and high doses of SY significantly reduced the levels of total cholesterol, triglycerides, and low-density lipoprotein cholesterol in the serum of hyperlipidemic mice and low-density lipoprotein cholesterol/high-density lipoprotein cholesterol ratio (P < 0.05), and the fatty liver of hyperlipidemic mice was significantly alleviated. SY had a protective effect on Chinese hamster ovary cells following amphotericin B injury (P < 0.01). SY exerts significant hypolipidemic effects and prevents fatty liver in a mechanism associated with inhibition of the biosynthesis of intracellular cholesterol.

Efficacy of Artemisia annua polysaccharides as an adjuvant to hepatitis C vaccination.

The traditional Chinese medicine Artemisia annua can prevent and treat hepatitis following an unclear mechanism. The aim of this study was to evaluate the effects of A. annua polysaccharides (AAP) on hepatitis C virus (HCV). A pcDNA3.1/NS3 expression vector was constructed. Ninety female BALB/c mice were randomly divided into six groups: high-dose AAP (1 mg/mL) + HCV/NS3 plasmid; middle-dose AAP (0.5 mg/mL) + HCV/NS3 plasmid; low-dose AAP (0.1 mg/mL) + HCV/NS3 plasmid; HCV/NS3 plasmid; high-dose AAP (1 mg/mL); normal saline control (N = 15). Except the control group and the high-dose AAP group, other groups were inoculated with 50 µg pcDNA3.1-HCV/NS3 plasmid. Serum antigenic-specific antibody was detected after the last immunization, and the levels of secreted IFN-γ and IL-4 were measured. pcDNA3.1/NS3 plasmid was successfully constructed, and the extracted product contained HCV/NS3 sequence. Compared with single inoculation with HCV/NS3 DNA vaccine, the specific antibody levels induced by middle-dose AAP plus HCV/NS3 DNA vaccine were significantly different in weeks 1, 3 and 5 (P < 0.05). However, there were no significant differences in the antibody levels induced by high-dose and low-dose AAP as adjuvant compared with those of single inoculation with DNA vaccine (P > 0.05). The level of serum IFN-γ secretion was significantly higher than that of IL-4 secretion. Compared with the single HCV/NS3 DNA vaccine group, AAP plus HCV/NS3 DNA vaccine groups had significant increased IFN-γ levels (P < 0.05), but the IL-4 levels were not significantly different among these groups (P > 0.05). AAP, as the adjuvant of HCV/NS3 DNA vaccine, can widely regulate the humoral immunity and cellular immune function of normal and cyclophosphamide-induced immunocompromised mice. AAP can promote IFN-γ secretion probably by inducing Th1-type cellular immune response.