[Blastic plasmacytoid dendritic cell tumor treated with DVT regimen: a case report and literature review].

Blastic plasmacytoid dendritic cell neoplasm (BPDCN) is a rare and highly aggressive hematological malignancy, there is no standard treatment and the prognosis is very poor. Affiliated Zhongshan Hospital of Dalian University report a case of 85-year-old BPDCN male patient treated with DVT regimen (decitabine combined with Venetoclax and thalidomide) and achieved complete remission. The patient with skin nodules and the pathology diagnosed BPDCN, the next generation sequencing of skin nodules showed mutations of IDH2 and ASXL1. DVT (decitabine combined with Venetoclax and thalidomide) has significant efficacy with rapid and deep remission for BPDCN, and the adverse effects is less, especially suitable for elderly patients who cannot tolerate intense chemotherapy.

[Determination of 14 volatile organic compounds in urine by headspace gas chromatography-mass spectrometry].

Objective: To establish a automatic headspace gas chromatography-mass spectrometry (HS/GC-MS) method for the determination of 14 volatile organic compounds in urine. Methods: In September 2022, 10 ml urine sample was taken into a 20 ml headspace bottle, balanced for 30 min at 65 ℃, and then detected by HS/GC-MS and quantified by external standard method. Results: The 14 volatile organic compounds showed good linearity at 0.2-8.0 µg/L and 0.1-4.0 µg/L, with correlation coefficients ranging from 0.9956-0.9999. The recoveries were 79.8%-113.1% with relative standard deviations 0.05%-0.27% when three different concentration levels were added. Detection limit was 0.03-0.05 µg/L. Conclusion: The method is simple and convenient, and the recovery and precision meet the requirements. It can be used for the determination of common volatile organic compounds in urine.

High temperature stress induced oxidative stress, gut inflammation and disordered metabolome and microbiome in tsinling lenok trout.

Tsinling lenok trout (Brachymystax lenok tsinlingensis Li) is a species of cold-water salmon that faces serious challenges due to global warming. High temperature stress has been found to damage the gut integrity of cold-water fish, impacting their growth and immunity. However, limited research exists on the causal relationship between gut microbial disturbance and metabolic dysfunction in cold-water fish induced by high temperature stress. To address this gap, we conducted a study to investigate the effects of high temperature stress (24 °C) on the gut tissue structure, antioxidant capacity, gut microorganisms, and metabolome reactions of tsinling lenok trout. Our analysis using 16 S rDNA gene sequencing revealed significant changes in the gut microbial composition and metabolic profile. Specifically, the abundance of Firmicutes and Gemmatimonadetes decreased significantly with increasing temperature, while the abundance of Bacteroidetes increased significantly. Metabolic analysis revealed a significant decrease in the abundance of glutathione, which is synthesized from glutamate and glycine, under high temperature stress. Additionally, there was a notable reduction in the levels of adenosine, inosine, xanthine, guanosine, and deoxyguanosine, which are essential for DNA/RNA synthesis. Conversely, there was a significant increase in the abundance of D-glucose 6 P. Furthermore, high temperature stress adversely affects intestinal structure and barrier function. Our findings provide valuable insights into the mechanism of high temperature stress in cold-water fish and serve as a foundation for future research aimed at mitigating the decline in production performance caused by such stress.

Locoregional radiotherapy improves survival outcomes in de novo metastatic nasopharyngeal carcinoma treated with chemoimmunotherapy.

BACKGROUND: We aimed to investigate the efficacy of locoregional radiotherapy (LRRT) in patients with de novo metastatic nasopharyngeal carcinoma (dmNPC) receiving chemotherapy combined with anti-programmed cell death receptor-1 monoclonal antibodies (anti-PD-1 mAbs) as first-line treatment and identify optimal candidates for LRRT. MATERIALS AND METHODS: We enrolled patients with dmNPC receiving platinum-based palliative chemotherapy and anti-PD-1 mAbs followed or not followed by LRRT from four centers. The endpoints were progression-free survival (PFS), objective response rate (ORR), and overall survival (OS). We used the inverse probability of treatment weighting (IPTW) to balance the baseline characteristics of the LRRT and non-LRRT groups to minimize selection bias before comparative analyses. Multivariate analyses were carried out using the Cox proportional hazards model. RESULTS: We included 163 patients with dmNPC (median follow-up: 22 months). The median PFS was 20 months, and the ORR was 92.0%; the median OS was not achieved. After IPTW adjustments, patients who received LRRT had a significant survival benefit over those not receiving LRRT (median PFS: 28 versus 15 months, P < 0.001). The Epstein-Barr virus DNA (EBV DNA) level after four to six cycles of anti-PD-1 mAbs [weighted hazard ratio (HR): 2.19, 95% confidence interval (CI) 1.22-3.92, P = 0.008] and LRRT (weighted HR: 0.58, 95% CI 0.34-0.99, P = 0.04) were independent prognostic factors. Patients with undetectable EBV DNA levels after four to six cycles of anti-PD-1 mAbs (early EBV DNA clearance) benefitted from LRRT (HR: 0.41, 95% CI 0.22-0.79, P = 0.008), whereas those with detectable levels did not (HR: 1.30, 95% CI 0.59-2.87, P = 0.51). CONCLUSIONS: Palliative chemotherapy combined with anti-PD-1 mAbs followed by LRRT was associated with improved PFS in patients with dmNPC, especially for patients with early EBV DNA clearance.

[Construction and evaluation of an artificial intelligence-based risk prediction model for death in patients with nasopharyngeal cancer].

OBJECTIVE: To screen the risk factors for death in patients with nasopharyngeal carcinoma (NPC) using artificial intelligence (AI) technology and establish a risk prediction model. METHODS: The clinical data of NPC patients obtained from SEER database (1973-2015). The patients were randomly divided into model building and verification group at a 7∶3 ratio. Based on the data in the model building group, R software was used to identify the risk factors for death in NPC patients using 4 AI algorithms, namely eXtreme Gradient Boosting (XGBoost), Decision Tree (DT), Least absolute shrinkage and selection operator (LASSO) and random forest (RF), and a risk prediction model was constructed based on the risk factor identified. The C-Index, decision curve analysis (DCA), receiver operating characteristic (ROC) curve and calibration curve (CC) were used for internal validation of the model; the data in the validation group and clinical data of 96 NPC patients (collected from First Affiliated Hospital of Bengbu Medical College) were used for internal and external validation of the model. RESULTS: The clinical data of a total of 2116 NPC patients were included (1484 in model building group and 632 in verification group). Risk factor screening showed that age, race, gender, stage M, stage T, and stage N were all risk factors of death in NPC patients. The risk prediction model for NPC-related death constructed based on these factors had a C-index of 0.76 for internal evaluation, an AUC of 0.74 and a net benefit rate of DCA of 9%-93%. The C-index of the model in internal verification was 0.740 with an AUC of 0.749 and a net benefit rate of DCA of 3%-89%, suggesting a high consistency of the two calibration curves. In external verification, the C-index of this model was 0.943 with a net benefit rate of DCA of 3%-97% and an AUC of 0.851, and the predicted value was consistent with the actual value. CONCLUSIONS: Gender, age, race and TNM stage are risk factors of death of NPC patients, and the risk prediction model based on these factors can accurately predict the risks of death in NPC patients.

Microbiology rapid on-site evaluation: a better method for Mucoid Pseudomonas Aeruginosa diagnosis in bronchiectasic patients.

OBJECTIVE: The aim of this study is to find an accurate and fast method to diagnose the pathogen of bronchiectasis. PATIENTS AND METHODS: Ten bronchiectasic patients diagnosed with Mucoid Pseudomonas Aeruginosa (MPA) in the past two years were analyzed. Accuracy and time were compared between microbiology rapid on-site evaluation (M-ROSE) and sputum bacterial culture. RESULTS: The accuracy rate of M-ROSE in the patients is 100% consistent with bacterial culture results. The average time of M-ROSE is about 4.3 min, which is over 1000 times shorter than that of sputum bacterial culture. CONCLUSIONS: M-ROSE may be a better method for etiological diagnosis of MPA.

[Prospective clinical study on the influence of implant crown and bridge prostheses characteristics on peri-implant disease].

Objective: To investigate the influence of different characteristics of implant-supported fixed dental prostheses on the incidence of peri-implant disease. Methods: Prospective cohort was established for patients who received implant-supported fixed dental prostheses at the Department of Prosthodontics, School of Stomatology, The Fourth Military Medical University between June 2014 and September 2015. Several patient-related factors, implant prostheses factors, and oral hygiene maintenance factors were collected. The Log-rank test was used to compare the peri-implant disease rates of various factors, and the Cox proportional hazards model was used to conduct multivariate study on single factor significant factors to analyze the impact on the incidence of peri-implant disease. Results: A total of 214 subjects and 351 implants were included in the cohort, the follow-up period was (11.0±3.5) months. Finally, 43.0% (92/214) of patients and 37.3% (131/351) of implants developed peri-implant diseases. The incidence of peri-implant mucositis and peri-implantitis was 41.1% (88/214) and 4.2% (9/214) at the subject level,and 34.2% (120/351) and 3.1% (11/351) at the implant level. Among the factors associated with the implant prosthesis, single factor Log-rank analysis showed that prostheses retention methods, proximal contact of the prostheses, occlusion situation were statistical significance (P<0.05). Multivariate analysis using Cox proportional hazards model showed that screw retention (hazard ratio=2.38, 95%CI: 1.42-3.99), proximal contact loss of the prostheses (hazard ratio=2.36, 95%CI: 1.31-4.27) were independent risk factors for peri-implant disease (P<0.05). Conclusions: Factors such as prostheses retention mode and proximal contact characteristics have important influence on the health status of the implant.

Exposure to traffic-related air pollution and changes in exhaled nitric oxide and DNA methylation in arginase and nitric oxide synthase in children with asthma.

BACKGROUND: Traffic-related air pollution (TRAP) has been associated with increased risk of airway inflammation in children with asthma. While epigenetic changes could potentially modulate TRAP-induced inflammatory responses, few studies have assessed the temporal pattern of exposure to TRAP, epigenetic changes and inflammation in children with asthma. Our goal was to test the time-lag patterns of personal exposure to TRAP, airway inflammation (measured as fractional exhaled nitric oxide, FeNO), and DNA methylation in the promoter regions of genes involved in nitric oxide synthesis among children with asthma. METHODS: We measured personal exposure to black carbon (BC) and FeNO for up to 30 days in a panel of children with asthma. We collected 90 buccal cell samples for DNA methylation analysis from 18 children (5 per child). Methylation in promoter regions of nitric oxide synthase (NOS1, NOS2A, NOS3) and arginase (ARG1, ARG2) was assessed by bisulfite pyrosequencing. Linear-mixed effect models were used to test the associations of BC at different lag periods, percent DNA methylation at each site and FeNO level. RESULTS: Exposure to BC was positively associated with FeNO, and negatively associated with DNA methylation in NOS3. We found strongest association between FeNO and BC at lag 0-6 h while strongest associations between methylation at positions 1 and 2 in NOS3 and BC were at lag 13-24 h and lag 0-24 h, respectively. The strengths of associations were attenuated at longer lag periods. No significant associations between exposure to TRAP and methylation levels in other NOS and ARG isoforms were observed. CONCLUSIONS: Exposure to TRAP was associated with higher levels of FeNO and lower levels of DNA methylation in the promoter regions of the NOS3 gene, indicating that DNA methylation of the NOS3 gene could be an important epigenetic mechanism in physiological responses to TRAP in children with asthma.

Postoperative radiotherapy option based on mediastinal lymph node reclassification for patients with pN2 non-small-cell lung cancer.

Background: In this research, we used the mediastinal lymph node reclassification proposed by the International Association for the Study of Lung Cancer (iaslc) to screen for patients with pathologic N2 (pN2) non-small-cell lung cancer (nsclc) who might benefit from postoperative radiotherapy (port). Methods: The study enrolled 440 patients with pN2 nsclc who received complete surgical resection and allocated them to one of three groups: N2a1 (single-station skip mediastinal lymph node metastasis), N2a2 (single-station non-skip mediastinal lymph node metastasis), and N2b (multi-station mediastinal lymph node metastasis). Rates of local recurrence at first recurrence in patients receiving and not receiving port were compared using the chi-square test. Overall (os) and disease-free survival (dfs) were then compared using Kaplan-Meier survival analysis with log-rank test. In addition, the factors potentially influencing os and dfs were analyzed using univariate and multivariate Cox regression. Results: The rate of local recurrence for the N2a2 and N2b groups was significantly lower in patients receiving port (p = 0.044 and p = 0.043 respectively). The log-rank test revealed that, for the N2a1 group, differences in os and dfs were not statistically significant between the patients who did and did not receive port (p = 0.304 and p = 0.197 respectively). For the N2a2 group, os and dfs were markedly superior in patients who received port compared with those who did not (p = 0.001 and p = 0.014 respectively). For the N2b group, os was evidently better in patients who received port compared with those who did not (p = 0.025), but no statistically significant difference in dfs was observed (p = 0.134). Multivariate regression analysis revealed that, in the N2a1 group, port was significantly associated with poor os [hazard ratio (hr): 2.618; 95% confidence interval (ci): 1.185 to 5.785; p = 0.017]; in the N2a2 group, port was associated with improved os (hr: 0.481; 95% ci: 0.314 to 0.736; p = 0.001) and dfs (hr: 0.685; 95% ci: 0.479 to 0.980; p = 0.039). Conclusions: For patients with pN2 nsclc who receive complete resection, port might be beneficial only for patients with single-station non-skip metastasis (N2a2). Patients with single-station skip metastasis (N2a1) and multi-station metastasis (N2b) might not currently benefit from port.

[The value of spectral CT-based radiomics in preoperative prediction of lymph node metastasis of advanced gastric cancer].

Objective: To investigate the spectral CT-based radiomics in predicting preoperatively the lymph node metastasis (LNM) of advanced gastric cancer. Methods: From January 2014 to October 2018, the spectral CT imaging and clinical data of 196 gastric adenocarcinoma patients confirmed by pathology in the First Affiliated Hospital of Zhengzhou University were retrospectively enrolled (training set and test set were randomly divided according to the ratio of 1∶1). These 196 patients include143 males and 53 females, aged from 28 to 81 years, with an average age of (59±11) years, and were divided into nodular metastasis group and non-metastasis group according to clinicopathological data. The spectral parameters were measured and calculated, and the CT-reported lymph node (LN) status from CT images were obtained. 273 radiomics features were extracted from the dual-phases CT images in different energy level (40, 65 and 100 keV) to build the radiomics signature respectively. Univariate analysis was used to compare the differences of spectral parameters and radiomics features between two groups, and then the significant indicators were put into multivariable logistic regression analysis to construct combined prediction model and radiomics nomogram. In addition, the performance of prediction model in training and test set were measured using the receiver operating characteristics (ROC) curves and were compared using DeLong test. Results: Both in training set and in test set, the iodine concentration (IC) of tumor in venous phase (VP) in nodular metastasis group were higher than that in non-metastasis group [training set: 22.98 (100 mg/L)>20.31 (100 mg/L), P=0.086; test set: 25.14 (100 mg/L)>21.07 (100 mg/L), P=0.009]. The CT-reported LN status showed significant differences between the two group (P<0.001, P=0.001). The radiomics signatures 40 keV-arterial phase, 65 keV-venous phase, IC-VP of tumor and CT-reported LN status were independent indicators for prediction of preoperative LNM of advanced gastric cancer in combined prediction model (P<0.05). The radiomics nomogram predicated LNM with an area under curve (AUC) and 95% confidence interval (CI) of 0.822 (0.739-0.906) in training set and 0.819(0.732-0.906) in test set, and there were no significant differences in AUC between two sets (P>0.05). Conclusions: The spectral CT-based radiomics can be used to quantitatively predict the LNM of advanced gastric cancer preoperatively.

Deep learning radiomic nomogram can predict the number of lymph node metastasis in locally advanced gastric cancer: an international multicenter study.

BACKGROUND: Preoperative evaluation of the number of lymph node metastasis (LNM) is the basis of individual treatment of locally advanced gastric cancer (LAGC). However, the routinely used preoperative determination method is not accurate enough. PATIENTS AND METHODS: We enrolled 730 LAGC patients from five centers in China and one center in Italy, and divided them into one primary cohort, three external validation cohorts, and one international validation cohort. A deep learning radiomic nomogram (DLRN) was built based on the images from multiphase computed tomography (CT) for preoperatively determining the number of LNM in LAGC. We comprehensively tested the DLRN and compared it with three state-of-the-art methods. Moreover, we investigated the value of the DLRN in survival analysis. RESULTS: The DLRN showed good discrimination of the number of LNM on all cohorts [overall C-indexes (95% confidence interval): 0.821 (0.785-0.858) in the primary cohort, 0.797 (0.771-0.823) in the external validation cohorts, and 0.822 (0.756-0.887) in the international validation cohort]. The nomogram performed significantly better than the routinely used clinical N stages, tumor size, and clinical model (P < 0.05). Besides, DLRN was significantly associated with the overall survival of LAGC patients (n = 271). CONCLUSION: A deep learning-based radiomic nomogram had good predictive value for LNM in LAGC. In staging-oriented treatment of gastric cancer, this preoperative nomogram could provide baseline information for individual treatment of LAGC.

Streamlined MRM method transfer between instruments assisted with HRMS matching and retention-time prediction.

Multiple reaction monitoring (MRM) mode using liquid-chromatography tandem mass spectrometry (e.g., LC-QqQ-MS/MS) has been extensively employed in the small molecule analysis with trace levels in complex samples owing to its high sensitivity. However, most of the reported MRM methods are developed using authentic standards, which are often costly yet not readily available. To address this question, a practical platform for the MRM method transfer between different LC-QqQ-MS/MS instruments, assisted by the high-resolution mass spectrometry (LC-HRMS) and retention time (RT) prediction, has been developed in this study. The reported platform can take advantage of both the high sensitivity of LC-MRM method and ion transition pairs from the previous publications. LC-HRMS can provide the accurate mass measurement of the compounds, though high-quality MS/MS fragments are usually difficult to obtain for chemicals at trace levels. Retention time matching and peaks matching between both instrumental platforms rule out isobaric candidates. With an additional retention time prediction filter from quantitative structure retention relationship (QSRR) model based on random forest feature selection (Pearson r2 = 0.63), identification of small molecules is achieved at a high confidence level without using authentic standards. The developed platform has been validated with robustness by examining spiked environmental chemicals in sludge water samples, biological urine, and cell extracts.

Development and validation of an individualized nomogram to identify occult peritoneal metastasis in patients with advanced gastric cancer.

BACKGROUND: Occult peritoneal metastasis (PM) in advanced gastric cancer (AGC) patients is highly possible to be missed on computed tomography (CT) images. Patients with occult PMs are subject to late detection or even improper surgical treatment. We therefore aimed to develop a radiomic nomogram to preoperatively identify occult PMs in AGC patients. PATIENTS AND METHODS: A total of 554 AGC patients from 4 centers were divided into 1 training, 1 internal validation, and 2 external validation cohorts. All patients' PM status was firstly diagnosed as negative by CT, but later confirmed by laparoscopy (PM-positive n = 122, PM-negative n = 432). Radiomic signatures reflecting phenotypes of the primary tumor (RS1) and peritoneum region (RS2) were built as predictors of PM from 266 quantitative image features. Individualized nomograms of PM status incorporating RS1, RS2, or clinical factors were developed and evaluated regarding prediction ability. RESULTS: RS1, RS2, and Lauren type were significant predictors of occult PM (all P < 0.05). A nomogram of these three factors demonstrated better diagnostic accuracy than the model with RS1, RS2, or clinical factors alone (all net reclassification improvement P < 0.05). The area under curve yielded was 0.958 [95% confidence interval (CI) 0.923-0.993], 0.941 (95% CI 0.904-0.977), 0.928 (95% CI 0.886-0.971), and 0.920 (95% CI 0.862-0.978) for the training, internal, and two external validation cohorts, respectively. Stratification analysis showed that this nomogram had potential generalization ability. CONCLUSION: CT phenotypes of both primary tumor and nearby peritoneum are significantly associated with occult PM status. A nomogram of these CT phenotypes and Lauren type has an excellent prediction ability of occult PM, and may have significant clinical implications on early detection of occult PM for AGC.

[Quantitative detection and clinical value of CD44 in patients with nonalcoholic fatty liver disease].

Objective: To quantitatively detect CD44 expression in patients with nonalcoholic fatty liver disease (NAFLD) for comparative analysis. Methods: Patients with chronic liver diseases accompanied with or without NAFLD, including chronic hepatitis B, cirrhosis and hepatocellular carcinoma after chronic hepatitis B, and healthy blood donors as normal controls who admitted to the Affiliated Hospital of Nantong University from May to October 2018 were selected. The proportion of CD44 positive cells was analyzed by flow cytometry. CD44 level was quantified by an enzyme-linked immunosorbent assay, and the biochemical indicators such as serum aspartate aminotransferase, alanine aminotransferase activity, total cholesterol and triglyceride were routinely analyzed. The cancerous and adjacent cancerous tissues of patients accompanied with or without NAFLD were collected by self-matching method and analyzed by immunoblotting and histochemistry and compared by CD44 integrated optical density. Image-Pro Plus version 6.0, Image J, GraphPad Prism 5.0, Photoshop, Microsoft Excel and IBM SPSS statistics 23 were used to analyze and draw pictures. An independent sample t-test was used to compare the differences between groups. Results: Patients accompanied with NAFLD had hepatocyte injury and dyslipidemia. NAFLD and chronic liver disease patients had significantly elevated serum CD44 levels than normal control group (P < 0.01). CD44 positive lymphocyte ratio was 78.19 % ± 16.33 % in NAFLD patients and 68.47% ± 20.91% in chronic hepatitis B group, which was higher than the control group (46.51% ± 20.52%). Chronic hepatitis B group with steatosis had significantly higher CD44 concentration (181.42 ± 49.36) ng/ml than chronic hepatitis B group (142.52 ± 53.87) ng/ml and normal control group (99.47 ± 15.23) ng/ml. CD44/GAPDH ratio in the liver cancer group (1.306 ± 0.614) was significantly higher than paracancerous group (0.477 ± 0.291) and the difference between the two groups was statistically significant (t = 3.451, P = 0.004). The integrated optical density of CD44 in the NAFLD-related liver cancer and paracancerous group were 25.721 ± 5.881 and 14.155 ± 4.001 and the difference between the groups was statistically significant (t = 14.544, P < 0.001). The pathological features of high expression of CD44 in patients with hepatocellular carcinoma were significantly correlated with HBV infection, tumor size, single/multi-center, and lymph node metastasis, degree of differentiation, TNM grade, Child-Pugh score, portal vein tumor thrombus and extrahepatic metastasis. HCC patients with NAFLD had significantly higher serum CD44 (234.62 ± 69.40) ng/ml than patients without NAFLD (186.49 ± 58.89) ng/ml (t = -3.191, P = 0.002), but there was no statistically significant difference in the clinicopathological characteristics between the high/low CD44 groups of HCC patients with NAFLD. Conclusion: The results suggest that CD44 is abnormally activated and its mechanism may play an important role in the progression of NAFLD.

[The anti-proliferative and anti-inflammatory mechanisms of JAK1 inhibitor SHR0302 versus Ruxolitinib in SET2 cell line and primary cells].

Objective: To explore the effects and molecular mechanism of the selective JAK1inhibitor SHR0302 and Ruxolitinib on myeloproliterative neoplasms (MPN) cell line SET2 and primary cells in vitro. Methods: Cell proliferation was detected by CCK8 kit. Colony forming experiment was conducted to evaluate erythroid burst colony formation unit (BFU-E) of primary cells from MPN patients. Multi-factor kits were used to detect six inflammatory cytokines. Phosphorylated proteins of Jak-Stat signaling pathway were tested by Western blot. Results: At different time points after treated with SHR0302 and Ruxolitinib, the inhibition of cell proliferation was dose dependent by both drugs (P<0.01) . The inhibitory rates of 2.5 µmol/L SHR0302 and 0.1 µmol/L Ruxolitinib on SET2 cells for 72 h were comparable, i.e. (59.94±0.60) % and (64.00±0.66) %, respectively, suggesting that the inhibitory effect of SHR0302 was weaker than that of Ruxolitinib. Similarly, both SHR0302 and Ruxolitinib inhibited BFU-E in primary marrow cells from MPN patients in a dose-dependent manner. SHR0302 1.0 µmol/L produced similar degree of inhibition compared to Ruxolitinib 0.2 µmol/L. Except IL-12, the expression of other 5 cytokines (IL-6, TNF-α, IL-1ß, IL-2, IL-8) was significantly inhibited by 1.6 µmol/L SHR0302 in SET2 cells at 24 h (P<0.01) , while Ruxolitinib 1.0 µmol/L had the same effect. Several phosphorylated molecules of Jak-Stat signaling pathway were significantly inhibited by SHR0302 in SET2 cells only for 3 h. P-stat1 (Tyr701) , p-stat3 (Tyr705) were down-regulated when treated with SHR0302 1.0 µmol/L (P<0.05) , p-jak1 (tyr1022/1023) and p-stat5 (Tyr694) were inhibited at 5.0 µmol/L (P<0.05) . Ruxolitinib significantly inhibited the downstream STAT protein at 0.1 µmol/L. Again, the inhibitory effect of SHR0302 on protein expression was weaker than that of Ruxolitinib. Conclusion: SHR0302 can effectively inhibit the proliferation of MPN cell line and patients' primary cells, as well as the expression of inflammatory factors. The molecular mechanism is possibly related to the down-regulation of phosphorylated proteins of Jak-Stat signaling pathway. Overall, the anti-proliferative and anti-inflammatory effects of SHR0302 are weaker than those of Ruxolitinib.

Oxidative stress mediated mitochondrial damage plays roles in pathogenesis of diabetic nephropathy rat.

OBJECTIVE: Diabetic nephropathy (DN) is considered as a complication of diabetes and accounts for about 40%. Reactive oxygen species (ROS) level continues to increase in DN. However, whether specific ROS levels can alleviate renal damage by improving mitochondrial function has not been investigated. MATERIALS AND METHODS: DN model was established by intraperitoneal injection of streptozotocin (STZ). The rats were divided into normal group, STZ model group, and N-acetylcysteine (NAC) group. Fasting blood glucose was tested to assess the modeling. The renal injury was evaluated by using hematoxylin-eosin (HE) staining and periodate-Schiff staining. Serum creatinine and 24 h urinary protein levels were determined by renal function detection kit. The levels of ROS and malondialdehyde (MDA) were assessed by the kit to evaluate the effects of oxidative stress and NAC in rats. The mitochondrial damage marker Cyto C level was detected by Western blot. RESULTS: Blood glucose, serum creatinine, and urinary protein levels were significantly increased in model group compared with the normal group (p<0.05). Blood glucose levels, serum creatinine, and urinary protein levels were markedly improved after the ROS and MDA levels reduced by NAC. Meanwhile, glomerular hypertrophy, mesangial matrix accumulation, and severe renal injury were observed in the model group. NAC administration markedly improved glomerular morphology and mesangial matrix aggregation. Cyto C expression in the model group increased significantly compared with normal control group (p<0.05), while NAC effectively inhibited Cyto C levels. CONCLUSIONS: Oxidative stress-mediated mitochondrial damage is an important part of the pathogenesis of DN. Inhibition of ROS production can be a potential target for DN treatment.

[Abnormal expression of angiopoietin-2 associated with invasion, metastasis and prognosis of lung cancer].

Objective: To investigate the relationship between abnormal angiopoietin-2 (Ang-2) expression and invasion/metastasis of lung cancer. Methods: Totally 122 cases of postoperative primary lung cancer tissues and their paracancerous tissues from Jan. 2009 to Dec. 2010 were collected from Affiliated Hospital of Nantong University and Ang-2 expression was analyzed by immunohistochemistry. At the cellular level, the protein and mRNA levels of Ang-2 in lung epithelial cell line Beas-2B and four lung cancer cell lines (SPCA-1, NCI-1650, A549 and NCI-H1975) were observed. The most effective Ang-2-shRNA for Ang-2 transcription was screened and transfected into A 549 lung cancer cells. The Ang-2 expression, Ang-2 gene transcription, cell proliferation, invasion/metastasis, and epithelial-mesenchymal transition (EMT) abilities of lung cancer cells were analyzed by Western blotting, fluorescent quantitative reverse transcriptase PCR, Cell Counting Kit-8 assay, and Transwell cell models for exploring the relationship between Ang-2 expression and invasion/metastasis of lung cancer. Results: The higher Ang-2 expression levels in lung cancerous tissues were closely related to tumor diameter (P=0.008), differentiating degree (P=0.033), TNM stage (P=0.025) and 5-year survival rate (P<0.001). According to the Kaplan-Meier survival curves, the 5-year survival rate of patients with higher expression levels of Ang-2 (16.1%) was significantly poorer than that of patients with lower Ang-2 (80.0%, P<0.001). Significant difference of 5-year survival rate was found in patients with different Ang-2 levels at TNM stage Ⅰ(P<0.001), but not at stage Ⅱ, Ⅲ and Ⅳ. Among Beas-2B and four lung cancer cell lines, the protein and mRNA levels of Ang-2 in A549 cells were the highest. After Ang-2-shRNA-1 plasmid successfully transfected into A549 cells, cell proliferation rate was significantly lower than that in the shRNA-negative or blank group at a time-dependent manner. The significant decrease of the invasion, migration and EMT abilities were also found in A549 cells after transfection of Ang-2 shRNA. Conclusion: Abnormal expression of Ang-2 is closely related to invasion, migration and prognosis of lung cancer, and interfering the activation of Ang-2 would be a novel molecular-targeted therapy for lung cancer.