[The value of CXorf67 and H3K27me3 for diagnosing germ cell tumors in central nervous system].

Objective: To investigate immunohistochemical patterns of CXorf67 and H3K27me3 proteins in central nervous system germ cell tumors (GCTs) and to assess their values in both diagnosis and differential diagnosis. Methods: A total of 370 cases of central nervous system GCTs were collected from 2013 to 2020 at Huashan Hospital of Fudan University, Shanghai, China. The expression of CXorf67, H3K27me3 and commonly-used GCT markers including OCT4, PLAP, CD117, D2-40, and CD30 by immunohistochemistry (EnVision method) was examined in different subtypes of central nervous system GCTs. The sensitivity and specificity of each marker were compared by contingency table and area under receiver operating characteristic (ROC) curve. Results: Of the 370 cases there were 282 males and 88 females with a mean age of 19 years and a median age of 17 years (range, 2-57 years). Among the GCTs with germinoma, the proportions of male patients and the patients with GCT located in sellar region were both higher than those of GCTs without germinoma (P<0.05), respectively. CXorf67 was present in the nuclei of germinoma and normal germ cells, but not in other subtypes of GCT. H3K27me3 was negative in germinoma, but positive in the nuclei of surrounding normal cells and GCTs other than germinoma. In the 283 GCTs with germinoma components, the expression rate of CXorf67 was 90.5% (256/283), but no cases were positive for H3K27me3. There was also an inverse correlation between them (r2=-0.831, P<0.01). The expression rates of PLAP, OCT4, CD117 and D2-40 were 81.2% (231/283), 89.4% (253/283), 73.9% (209/283) and 88.3% (250/283), respectively. In 63 mixed GCTs with germinoma components, the expression rate of CXorf67 was 84.1% (53/63), while all cases were negative for H3K27me3. The expression rates of PLAP, OCT4, CD117 and D2-40 were 79.4% (50/63), 79.4% (50/63), 66.7% (42/63) and 87.3% (55/63), respectively. The 6 markers with largest area under ROC curve in ranking order were H3K27me3, CXorf67, D2-40, OCT4, PLAP and CD117 (P<0.05). Conclusions: CXorf67 and H3K27me3 have high sensitivity and high specificity in diagnosing germinoma. There is a significant inverse correlation between them. Therefore, they can both be used as new specific immunohistochemical markers for the diagnosis of GCTs.

Prognostic potentials of miRNA-19a-3p and PDCD5 in nasopharynx carcinoma.

OBJECTIVE: To determine expressions of MicroRNA-19a-3p (miRNA-19a-3p) and PDCD5 in nasopharyngeal carcinoma (NPC) tissues, and their prognostic potentials in NPC. PATIENTS AND METHODS: Expressions of miRNA-19a-3p and PDCD5 in NPC tissues and controls were determined by quantitative Real Time-Polymerase Chain Reaction (qRT-PCR). The correlation between expressions of miRNA-19a-3p and PDCD5 in NPC was evaluated by Pearson correlation test. Furthermore, potential influences of miRNA-19a-3p and PDCD5 on clinical features of NPC patients were assessed. Through 5-year follow-up, survival analysis in NPC patients was conducted by Kaplan-Meier method. Finally, factors influencing prognosis of NPC were determined using the Cox regression model. RESULTS: MiRNA-19a-3p was upregulated and PDCD5 was downregulated in NPC tissues. Pearson correlation test uncovered a negative correlation between expression levels of miRNA-19a-3p and PDCD5 in NPC tissues. MiRNA-19a-3p level was correlated with N classification and clinical stage in NPC patients, while PDCD5 level was correlated with T classification, pathological grade and clinical stage. Survival analysis showed poor prognosis in NPC patients expressing high level of miRNA-19a-3p or low level of PDCD5. Cox regression analysis illustrated that N2+3 classification, clinical stage III+IV, high level of miRNA-19a-3p and low level of PDCD5 were independent risk factors for the prognosis of NPC. CONCLUSIONS: MiRNA-19a-3p is upregulated and PDCD5 is downregulated in NPC tissues. High level of miRNA-19a-3p and low level of PDCD5 are unfavorable for the prognosis of NPC.

[Protective effect and mechanism of mild hypothermia on liver injury after cardiopulmonary resuscitation in pigs].

Objective: To investigate the effect of mild hypothermia therapy on liver after cardiopulmonary resuscitation. Methods: Thirty-three inbred Chinese Wuzhishan (WZS) minipigs, weighted (28±2) kg, were used to establish a ventricular fibrillation model. And 30 animals survived after cardiopulmonary resuscitation reached basic life support. The surviving animals were randomly divided into two groups: mild hypothermia group (group M, n=15) and conventional treatment group (group C, n=15). All the animals were observed for 24 hours. Blood samples were extracted at baseline, 0.5, 1, 2, 4, 6, 12 and 24 h after successful resuscitation. The concentrations of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) were detected at the time points. The enzyme-linked immunosorbent assay (ELISA) was used to detect the concentrations of interleukin-6 (IL-6) and tumour necrosis factor-alpha (TNF-α). The data were compared between the two groups, LSD test was used when the variance was homogeneous, and Tamhane T2 test was used when the variance was uneven. Results: Eleven pigs (73.3%) in the group M and 8(53.3%) in the group C survived at 24 h after successful resuscitation, with no statistically significant difference between the two groups (χ(2)=1.229, P=0.225). After successful resuscitation, the AST, ALT increased in both group but less in M group (all P<0.05). After successful resuscitation, the concentrations of TFN-α and IL-6 in the blood increased in both groups, reached the peak at 4h, and then decreased gradually. The concentrations of TFN-α in group M were lower than those in group C at 0.5, 2, 4 and 6 h after successful resuscitation (t=0.01, 0.01, 0.87, 0.86, all P<0.05). The concentrations of IL-6 in the group M were lower than those in group C at 0.5, 1, 2 and 4 h after successful resuscitation (t=0.23, 0.78, 0.11, 0.80, all P<0.05). Conclusions: After successful resuscitation, the release of inflammatory mediators, such as TNF-α and IL-6, and cell apoptosis may involve in liver ischemia reperfusion injury. After successful resuscitation, the liver undergoes ischemia-reperfusion injury, which may be related to the release of inflammatory mediators such as TNF-α and IL-6. Mild hypothermia therapy can prevent the release of TNF-α, IL-6 to reduce the degree of liver damage after resuscitation.

Circ_CHFR expedites cell growth, migration and inflammation in ox-LDL-treated human vascular smooth muscle cells via the miR-214-3p/Wnt3/ß-catenin pathway.

OBJECTIVE: Atherosclerosis (AS) is a representative inflammatory vascular disease. This study explored the molecular pathogenesis of AS based on circular RNA (circRNA), the checkpoint with forkhead-associated and ring-finger domains (circ_CHFR). PATIENTS AND METHODS: The cell model of AS in vitro was established by stimulating human vascular smooth muscle cells (VSMCs) with oxidized low-density lipoprotein (ox-LDL). The RNA expression was measured by quantitative Real-time polymerase chain reaction (qRT-PCR). Cell viability and colony formation ability were separately evaluated using 3-(4, 5-dimethylthiazol-2-y1)-2, 5-diphenyl tetrazolium bromide (MTT) and colony formation assay. Cell migration was assessed via the transwell assay. The inflammation injury was analyzed by enzyme-linked immunosorbent assay (ELISA). Associated proteins were determined through Western blot. The combination of hypothetic targets was ascertained using Dual-Luciferase reporter assay. RESULTS: Circ_CHFR was up-regulated in AS serums and ox-LDL-stimulated VSMCs. Circ_CHFR depletion weakened the ox-LDL-induced promotion of cell growth, migration and inflammation in VSMCs. Circ_CHFR positively regulated Wnt3 expression and the downregulation of Wnt3 abrogated the ox-LDL-triggered injuries in VSMCs. Circ_CHFR functioned as the sponge of microRNA-214-3p (miR-214-3p) and miR-214-3p targeted Wnt3. Circ_CHFR regulated cell growth, migration and inflammation via regulating the expression of Wnt3 as a competitive endogenous RNA (ceRNA) of miR-214 in ox-LDL-treated VSMCs. Circ_CHFR/miR-214-3p axis mediated the Wnt3/ß-catenin signal pathway. CONCLUSIONS: Circ_CHFR contributed to the progression of AS through the miR-214-3p/Wnt3/ß-catenin signals, which illuminated the molecular mechanism of AS and suggested circ_CHFR might be an index for AS treatment.

[Study on the relationship between preoperative evaluation, operation and prognosis of microvascular decompression for the treatment of hemifacial spasm].

Objective: To investigate the relationship between preoperative evaluation, surgery and prognosis of microvascular decompression (MVD) for the treatment of hemifacial spasm (HFS). Methods: The clinical data of 128 HFS patients treated with MVD in the department of neurosurgery of Taizhou Hospital of Zhejiang Province were retrospectively analyzed. According to the SMC grading system, the patients were divided into general spasm group and severe spasm group, and the clinical characteristics, offending vessel, prognosis and surgical complications of the two groups were compared. Results: In the general spasm group,the age at MVD was (48.6±10.6) years, the disease duration was (4.2±3.3) years;while in the severe spasm group,the age at MVD was (51.8±9.9) years, the disease duration was (8.1±4.5) years;the differences of age and disease duration between the two groups were statistically significant (P<0.05).In the general spasm group, there were 41 cases in which the offending vessel were AICA, 21 cases were PICA, 1 case was VA, 63 cases were single offending vessel, and 7 cases were multiple offending vessels. In the severe spasm group, there were 29 cases in which the offending vessel were AICA, 13 cases were PICA, 2 cases were VA, the total of 44 cases were single offending vessel and 14 cases were multiple offending vessels.There was a significant difference in the proportion of multiple offending vessels in the two groups, and the difference was statistically significant (P<0.05).Patients in the two groups were followed up for 12 to 32 months after surgery, and the difference in effective rate and recurrence rate was not statistically significant (P>0.05).Some kinds of postoperative complications were different between the two groups, the incidence of postoperative delayed facial paralysis was statistically significant (P<0.05), and the other complications were not statistically significant (P>0.05). Conclusion: Compared with the general spasm group, the patients in the severe spasm group were older, with longer disease duration, higher probability of multiple offending vessels and higher incidence of postoperative delayed facial paralysis. Therefore, preoperative SMC grading is helpful for the evaluation and prediction of intraoperative and postoperative conditions, which is worthy of wide clinical application.

[Clinical efficacy of different vascular reconstruction on adult moyamoya disease].

Objective: To investigate the clinical efficacy of superficial temporal artery -middle cerebral artery combined with encephalo-duro-arterio-myo-synangiosis (STA-MCA+EDAMS) and encephalo-duro-arterio-myo-synangiosis (EDAMS) in the treatment of adult moyamoya disease. Methods: The clinical data of 47 adult patients with moyamoya disease who received vascular reconstruction in the Department of Neurosurgery of Taizhou Hospital of Zhejiang Province from January 2014 to January 2018 were retrospectively analyzed. Among them, 21 patients received EDAMS alone (EDAMS group, 14 patients with hemorrhagic moyamoya disease, 7 patients with ischemic moyamoya disease), 26 patients received STA-MCA combined with EDAMS (STA-MCA+EDAMS group, 17 patients with hemorrhagic moyamoya disease, 9 patients with ischemic moyamoya disease). Cerebral hemodynamics at 1 day before surgery and 3 and 6 months after surgery were compared. The clinical efficacy and postoperative complications of the two methods were compared at 3 and 6 months postoperatively in hemorrhagic and ischemic types. Results: For hemorrhagic moyamoya disease, the remission rate (94.1%) at 6 months after surgery in the STA-MCA + EDAMS group was higher than that in the EDAMS group (57.1%), and the difference was statistically significant (P<0.05). The CBF and CBV in the STA-MCA+EDAMS group were higher than those in the EDAMS group at 3 and 6 months after operation, and the MTT and TPP were lower than those in the EDAMS group, but there was no significant difference between the two groups (all P>0.05). For hemorrhagic moyamoya disease and ischemic moyamoya disease, the total incidence of postoperative complications of the two surgical methods was different, but the difference was not statistically significant (both P>0.05). Conclusion: Superficial temporal artery -middle cerebral artery combined with encephalo-duro- arterio-myo-synangiosis (STA-MCA+EDAMS) and encephalo-duro-arterio-myo-synangiosis (EDAMS) can significantly improve neurological function and cerebral hemodynamics in adult moyamoya disease patients with high safety.

MicroRNA-384 downregulates SETD8 expression to suppress cell growth and metastasis in osteosarcoma cells.

OBJECTIVE: MiR-384 was reported to be downregulated and functioned as a tumor suppressor in several cancers. However, the expression and function of miR-384 in osteosarcoma (OS) have not been investigated. In the present study, we aimed to analyze the effect and mechanism of miR-384 in the progression of OS. PATIENTS AND METHODS: Quantitative Real-time polymerase chain reaction (qRT-PCR) was used to determine the expression of miR-384 in OS tissues and cells. MTT assay, colony formation analysis, Transwell assays were performed to analyze the role of miR-384 in human OS cells. Western blotting was applied to analyze the expression of SETD8, and the luciferase reporter assay was used to assess the target gene of miR-384 in OS cells. RESULTS: We found that miR-384 was significantly lowly expressed in OS tissues and OS cell lines compared with the adjacent noncancerous tissues and normal bone cell lines, respectively. Further functional analysis indicated that up-regulation of miR-384 significantly inhibited OS cells proliferation, migration, and invasion, but down-regulation of miR-384 had the opposite effects on OS cells in vitro. Moreover, SETD8 was identified as the potential target of miR-384 using dual luciferase assay, qRT-PCR and Western blot. Finally, we observed that upregulation of SETD8 reversed the effects of overexpressing of miR-384 on the proliferation, migration, and invasion of OS. CONCLUSIONS: Our data provided the first evidence which supported the function of miR-384 as a tumor suppressor in OS by targeting SETD8.

Clinical significance of long non-coding RNA EWSAT1 as a novel prognostic biomarker in osteosarcoma.

OBJECTIVE: Long non-coding RNA EWSAT1 (EWSAT1) has been identified as a tumor promoter in several tumors, but its prognostic values in osteosarcoma have not been reported. The purpose of this study was to explore the association between EWSAT1 expression and prognosis of osteosarcoma patients. PATIENTS AND METHODS: EWSAT1 levels were examined in 176 osteosarcoma tissues and matched normal bone tissues by qRT-PCR analysis. The associations of EWSAT1 expression with clinicopathologic variables were analyzed. The survival curves were calculated by the Kaplan-Meier method. The Cox proportional hazards regression model was used to identify independent prognostic factors with independent prognostic for overall survival (OS) and disease-free survival (DFS). RESULTS: We found that EWSAT1 levels were significantly higher in osteosarcoma tissues compared with matched non-cancerous tissues (p<0.01). The level of EWSAT1 expression was significantly associated with clinical stage (p=0.001) and distant metastasis (p=0.011). Then, Kaplan-Meier analysis showed that high EWSAT1 expression level was associated with poorer OS (p=0.0007) and DFS (p=0.0010). Furthermore, Cox multivariate analyses demonstrated that EWSAT1 expression was an independent prognostic factor for both OS (p<0.001) and DFS (p=0.001) in osteosarcoma patients CONCLUSIONS: Increased EWSAT1 expression was associated with poor outcomes in osteosarcoma patients, and EWSAT1 could serve as a potential unfavorable prognostic biomarker.

[Effect of the ischemic post-conditioning on the prevention of the cardio-renal damage in patients with acute ST-segment elevation myocardial infarction after primary percutaneous coronary intervention].

Objective: To evaluate the effect of the ischemic post-conditioning (IPC) on the prevention of the cardio-renal damage in patients with acute ST-segment elevation myocardial infarction (STEMI) after primary percutaneous coronary intervention (PPCI). Methods: A total of 251 consecutive STEMI patients underwent PPCI in the heart center of Tianjin Third Central Hospital from January 2012 to June 2014 were enrolled in this prospective, randomized, control, single-blinded, clinical registry study. Patients were randomly divided into IPC group (123 cases) and control group (128 cases) with random number table. Patients in IPC group underwent three times of inflation/deflation with low inflation pressure using a balloon catheter within one minute after culprit vessel blood recovery, and then treated by PPCI. Patients in control group received PPCI procedure directly. The basic clinical characteristics, incidence of reperfusion arrhythmia during the procedure, the rate of electrocardiogram ST-segment decline, peak value of myocardial necrosis markers, incidence of contrast induced acute kidney injury(CI-AKI), and one-year major adverse cardiovascular events(MACE) which including myocardial infarction again, malignant arrhythmia, rehospitalization for heart failure, repeat revascularization, stroke, and death after the procedure were analyzed between the two groups. Results: The age of IPC group and control group were comparable((61.2±12.6) vs. (64.2±12.1) years old, P=0.768). The incidence of reperfusion arrhythmia during the procedure was significantly lower in the IPC group than in the control group(42.28% (52/123) vs. 57.03% (73/128), P=0.023). The rate of electrocardiogram ST-segment decline immediately after the procedure was significantly higher in the IPC group than in the control group (77.24% (95/123) vs. 64.84% (83/128), P=0.037). The peak value of myocardial necrosis markers after the procedure were significantly lower in the IPC group than in the control group(creatine kinase: 1 257 (682, 2 202) U/L vs. 1 737(794, 2 816)U/L, P=0.029; creatine kinase-MB: 123(75, 218)U/L vs.165(95, 288)U/L, P=0.010). The rate of CI-AKI after the procedure was significantly lower in the IPC group than in the control group(5.69%(7/123) vs. 14.06%(18/128), P=0.034). The rate of the one-year MACE was significantly lower in the IPC group than in the control group(7.32%(9/123) vs. 15.63% (20/128), P=0.040). Conclusion: The IPC strategy performed eight before PPCI can reduce myocardial ischemia- reperfusion injury, decline the rates of CI-AKI and one-year MACE significantly in STEMI patients, thus has a significant protective effect on heart and kidney in STEMI patients. Clinical Trial Registration Chinese Clinical Trials Registry, ChiCTR-ICR-15006590.

Association between the XPG gene Asp1104His polymorphism and lung cancer risk.

It has been suggested that the xeroderma pigmentosum complementation group G (XPG) gene Asp1104His polymorphism is linked to susceptibility to lung cancer. However, the results from the published studies are contradictory rather than conclusive. With this meta-analysis, we aimed to achieve a better understanding of the effects of the XPG gene Asp1104His polymorphism on lung cancer risk. We identified six eligible studies from five publications that included a total of 2293 lung cancer patients and 2586 controls. There was a significant association between the XPG gene Asp1104His polymorphism and lung cancer (His/His vs Asp/Asp: OR = 1.24, 95%CI = 1.04-1.48; Asp/His vs Asp/Asp: OR = 1.17, 95%CI = 1.03-1.34; the dominant model: OR = 1.18, 95%CI = 1.04-1.33; the recessive model: OR = 1.10, 95%CI = 0.94-1.28). In a subgroup analysis by nationality, we found a significant association between the XPG gene Asp1104His polymorphism and lung cancer risk in Asians. No publication bias was found in this study. The results from this meta-analysis indicate that the XPG gene Asp1104His polymorphism is associated with lung cancer risk, especially in Asians.

Nemo-like kinase is critical for p53 stabilization and function in response to DNA damage.

The DNA damage response (DDR) acts as a protective mechanism for maintaining cell homeostasis. Nemo-like kinase (NLK) is a serine/threonine-protein kinase that has an important role in many pathways; however, its function in the DDR has not yet been defined. In our study, NLK-deficient HCT116 cells were found to be resistant to etoposide-induced cell death. We demonstrated that NLK is required for p53 activation in response to DNA damage. Remarkably, mechanistic studies revealed that NLK interacts with p53 and stabilizes p53 by blocking MDM2-mediated p53 ubiquitination and degradation. Furthermore, NLK enhances p53 activity and affects expression downstream of p53. Interestingly, these functions of NLK are not related to its kinase activity. Consistent with these results, NLK-deficient cells have a resistance effect on DNA damage. Therefore, these findings emphasize that NLK is a novel factor in DDR mechanisms.

Separation of oil from oily wastewater by modified resin.

A new type of packing material, polystyrene resin modified by cetyltrimethyl-ammonium bromide (R-CTAB), was developed for separation of emulsified oil wastewater. The unique separation mechanism of the modified resin is due to the hydrogen bond formation between hydrocarbon molecules and the free hydrophilic part of the fixed surfactant. It changed the zeta potential of oil droplets and made them coalescing easier, which provide an optional choice without any chemicals added into the wastewater. A column of R-CTAB was investigated for its performance in separating emulsified oil droplets in oily wastewater. The effects of liquid space velocity and influent oil concentration were studied in particular. The results showed that R-CTAB column was more efficient than the un-modified resin column. Under the optimum experimental conditions, more than 85% of oil was removed by using R-CTAB column that at least 10% higher than the un-modified resin column.