Jianjun Xu

Tissue spray ionization mass spectrometry for rapid recognition of human lung squamous cell carcinoma

Tissue spray ionization mass spectrometry (TSI-MS) directly on small tissue samples has been shown to provide highly specific molecular information. In this study, we apply this method to the analysis of 38 pairs of human lung squamous cell carcinoma tissue (cancer) and adjacent normal lung tissue (normal). The main components of pulmonary surfactants, dipalmitoyl phosphatidylcholine (DPPC, m/z 757.47), phosphatidylcholine (POPC, m/z 782.52), oleoyl phosphatidylcholine (DOPC, m/z 808.49), and arachidonic acid stearoyl phosphatidylcholine (SAPC, m/z 832.43), were identified using high-resolution tandem mass spectrometry. Monte Carlo sampling partial least squares linear discriminant analysis (PLS-LDA) was used to distinguish full-mass-range mass spectra of cancer samples from the mass spectra of normal tissues. With 5 principal components and 30 – 40 Monte Carlo samplings, the accuracy of cancer identification in matched tissue samples reached 94.42%. Classification of a tissue sample required less than 1 min, which is much faster than the analysis of frozen sections. The rapid, in situ diagnosis with minimal sample consumption provided by TSI-MS is advantageous for surgeons. TSI-MS allows them to make more informed decisions during surgery.

D2 dopamine receptor antagonist raclopride induces non‐canonical autophagy in cardiac myocytes

Cell death by autophagy is an important means of maintaining cellular homeostasis in adult cardiac myocytes. Autophagy was previously shown to exert a cardioprotective effect, suggesting that modulation of autophagy pathways is a potential therapeutic strategy in the treatment of heart disease. Although dopamine is known to induce autophagy in neuroblastoma cells, the underlying mechanism and the types of dopamine receptors involved in this process remain unclear. In this study, we used various dopamine receptor antagonists and agonists to identify the specific dopamine receptor that mediates induction of autophagy. We evaluated autophagy induction by the expression of autophagy markers in neonatal rat ventricular cardiac myocytes. We evaluated intracellular calcium levels using Fluo‐3/AM and demonstrated autophagy‐induced morphological changes in cardiac myocytes using electron microscopy. We also examined the pathway for dopamine‐induced autophagy using RNAi‐mediated gene knockdown. Raclopride, the well‐documented D2 receptor antagonist, significantly upregulated autophagy in cardiac myocytes via an mTOR‐independent pathway. There was no difference in intracellular calcium levels between raclopride‐treated cells and untreated cells. siRNA‐mediated knockdown of Rab9 resulted in decreased expression of autophagy markers in raclopride‐treated cells. Interestingly, siRNA‐mediated knockdown of Atg7 resulted in a significant increase in Rab9 levels in raclopride‐treated cells, suggesting that blocking the classical autophagy pathway results in activation of an alternative pathway. Our study suggests that (1) the D2 dopamine receptor plays a role in autophagy and (2) raclopride mediated a non‐canonical autophagy pathway in cardiac myocytes via Rab9. J. Cell. Biochem. 114: 103–110, 2012.

High level of BRD4 promotes non-small cell lung cancer progression.

Bromodomain containing protein 4 (BRD4), a member of the bromodomain and extra terminal domain (BET) protein family, has been shown to play important roles in tumor progression. However, its role in non-small cell lung cancer (NSCLC) is still largely unknown. Here, we found that BRD4 expression was significantly upregulated in NSCLC tissues and NSCLC cell lines with higher invasion and metastasis potentials. Suppression of BRD4 expression in NSCLC cell lines impaired cell invasion, inhibited cell proliferation, and accelerated cell apoptosis. Clinically, we observed that the BRD4 level was significantly related to histological type, lymph node metastasis, tumor stage and differentiation. More importantly, high level of BRD4 was closely correlated with the poor prognosis of NSCLC patients. Therefore, our study suggests that BRD4 is one of the major contributors to the invasion-prone phenotype of NSCLC, and a potential therapeutic target of NSCLC.

Correlation between single nucleotide polymorphisms in CYP4F2 and warfarin dosing in chinese valve replacement patients

BackgroundIndividuals with implanted mechanical valve prostheses require lifelong anticoagulation therapy with warfarin. The narrow therapeutic index of warfarin makes it difficult to dose and maintain proper anticoagulation. A number of single nucleotide polymorphisms (SNPs) affecting vitamin K or warfarin metabolism have been shown to affect warfarin dosing. Our aim was to study the effect of the CYP4F2 rs2108622-1347 (C > T) variant on warfarin dosing in Chinese patients.MethodsWe studied 352 patients after heart valve replacement surgery. Warfarin dosing for patients was adjusted to achieve 1.8 ≤ INR ≤ 2.5. We determined the presence of SNPs in CYP4F2 in these patients and investigated their association with warfarin dosing.ResultsWe found the frequency of the CYP4F2 rs2108622 C allele was 79.5% and T-allele frequency was 20.5%. The warfarin dose requirement for CC individuals was significantly lower than that for CT or TT individuals (P < 0.05). TT-homozygous individuals required a 0.56 mg/day higher dose of warfarin than their CC counterparts.ConclusionsThis study demonstrates that CYP4F2 rs2108622 significantly affects the warfarin dose requirement to achieve adequate anticoagulant activity in Chinese individuals. Genotyping of this SNP may allow clinicians to determine the initiation dose for patients following valve-replacement surgery in China.

A systematic review and meta-analysis of T2, T3 or T4, to evaluate the best denervation level for palmar hyperhidrosis

We systematically reviewed and compared the clinical outcomes of thoracoscopic sympathectomy (TS) at different denervation levels for palmar hyperhidrosis. We searched PubMed, Ovid MEDLINE, EMBASE, Web of Science, ScienceDirect, the Cochrane Library, Scopus and Google Scholar for relevant studies published during 1990–2016. Symptom resolution, patient satisfaction, compensatory sweating (CS), recurrence, dry hands and gustatory sweating were assessed. We selected 13 studies from 2228 for the final analysis. A comparison of T2 vs. T3 TS revealed that T3 TS reduced the risk of CS (95% confidence interval [CI]: 1.36–3.19, p = 0.0007) and moderate-to-severe CS (95% CI: 2.14–7.87, p < 0.0001). No significant differences were found in patient satisfaction, symptom resolution, and incidence of dry hands and gustatory sweating. A comparison of T3 vs. T4 TS revealed that T4 TS reduced the risk of CS (95% CI: 2.87–9.53, p < 0.00001), moderate-to-severe CS (95% CI: 2.54–5.83, p < 0.00001), dry hands (95% CI: 4.07–18.13, p < 0.00001) and gustatory sweating (95% CI: 1.53–7.32, p < 0.003), and improved patient satisfaction. No significant differences were found in symptom resolution and recurrence. T4 TS appears to be more useful than T3 or T2 TS for PH.

Prognostic roles of mRNA expression of notch receptors in non-small cell lung cancer

Notch signalling is aberrantly activated in human non-small cell lung cancer (NSCLC). Nevertheless, the prognostic roles of mRNA expression of four Notch receptors in NSCLC patients remain elusive. In this report, we reported the prognostic roles of Notch receptors in a total of 1,926 NSCLC patients through “The Kaplan-Meier plotter” (KM plotter) database which is capable to assess the effect of 22,277 genes on survival of NSCLC patients. We found that mRNA high expression level of Notch1 was associated with better overall survival (OS) for all NSCLC patients, hazard ratio (HR) 0.78 (0.69-0.89), p=0.00019, better OS in adenocarcinoma (Ade) patients, HR 0.59 (0.46-0.75), p=1.5e-05, as well as in squamous cell carcinoma (SCC) patients, HR 0.78 (0.62-0.99), p=0.044. mRNA high expression levels of Notch2 and Notch3 were associated with worsen OS for all NSCLC patients, as well as in Ade, but not in SCC patients. mRNA high expression level of Notch4 was not found to be associated with to OS for all NSCLC patients. In addition, mRNA high expression levels of Notch2, Notch3, but Notch4 are significantly associated with the NSCLC patients who have different smoking status. Our results indicate that mRNA expression of Notch receptors may have distinct prognostic values in NSCLC patients. These results will benefit for developing tools to accurately predict the prognosis of NSCLC patients.

Altered phosphatidylcholines expression in sputum for diagnosis of non-small cell lung cancer

Non–small cell lung cancer (NSCLC) is a leading cause of cancer mortality worldwide, and early diagnosis needs to be improved. We examined whether neutral desorption extractive electrospray ionization mass spectrometry (ND-EESI-MS) could be used to detect sputum lipids expression changes to enable earlier diagnosis. Overall, 167 NSCLC patients and 140 controls were enrolled. The main peaks in the sputum of patients with NSCLC patients differed from controls (83.3% of total variability), and the signals were not associated with pathological type, TNM stage or smoking history. The relative abundance of peaks at m/z734, m/ z756, m/z772, m/z782, m/z798 and m/z803 reliably distinguished NSCLC sputum from control. Collision-induced dissociation confirmed that m/z734, m/z756, and m/z772 represented [DPPC + H]+, [DPPC + Na]+, and [DPPC + K]+, respectively, and m/z782, m/z798, and m/z803 represented sphingomyelin, phosphatidylglycerol, and phosphatidylglycerolphosphate, respectively. The relative abundance of DPPC was clearly lower in NSCLC sputum than in control, and the relative abundances of phosphatidylglycerol and phosphatidylglycerolphosphate were higher in NSCLC sputum than in control. The detection of changes in sputum lipids with ND-EESI-MS may be a noninvasive, radiation-free, relatively inexpensive, repeatable, and efficient method for diagnosis of NSCLC.

The benefit and risk of nivolumab in non‐small‐cell lung cancer: a single‐arm meta‐analysis of noncomparative clinical studies and randomized controlled trials

Nivolumab is a programmed cell death 1 (PD‐1) receptor inhibitor antibody that enhances immune system antitumor activity. Although it is used for treating advanced non‐small‐cell lung cancer (NSCLC), its actual efficacy has not been determined. We searched PubMed, the Cochrane Library, Embase, MEDLINE, and Web of Science for related noncomparative clinical studies and randomized controlled trials (RCTs) to assess nivolumab benefit and risk in NSCLC. The main outcomes were objective response rate (ORR), 1‐year overall survival rate (1‐yOS rate), and progression‐free survival rate at 24 weeks (PFS at 24 weeks rate), any‐grade adverse effects rate (any‐grade AEs%), and grade 3–4 AE rate (grade 3–4 AEs%). Relative risk (RR) was used to compare ORR in patients with positive and negative programmed cell death ligand 1 (PD‐L1) expression. Random‐effects models were used to determine pooled effect size and two‐sided 95% confidence intervals (95% CI). We included 20 studies (17 noncomparative open‐label cohort studies, three RCTs) involving 3404 patients in our meta‐analysis. The modified nivolumab ORR was 18% (95% CI: 15–20%), the 1‐yOS rate was 45% (95% CI: 40–50%), PFS at 24 weeks rate was 42% (95% CI: 37–48%), any‐grade AEs% was 61% (95% CI: 50–73%), and grade 3–4 AEs% was 12% (95% CI: 9–16%). PD‐L1 expression was related with the nivolumab ORR. Nivolumab potentially causes ongoing response, long‐term PFS, and reduced treatment‐related AEs. PD‐L1 expression predicts the outcome of nivolumab immunotherapy. More high‐quality and well‐designed RCTs with large sample sizes are warranted to prove our findings.

Ring finger protein 38 promote non-small cell lung cancer progression by endowing cell EMT phenotype

Objectives: Ring finger protein 38 (RNF38), as an E3 ubiquitin ligase, plays an essential role in multiple biological processes by controlling cell apoptosis, cell cycle and DNA repair, and resides in chromosome 9 (9p13) which is involvement in cancer pathogenesis including lung cancer. However, its function in tumorigenesis remains unclear. Hence, this study set out to investigate the biological function and clinical implications of RNF38 in non-small cell lung cancer (NSCLC). Materials and Methods: Immunohistochemistry, quantitative real-time polymerase chain reaction (qRT-PCR) and western blot were used to detect RNF38 protein and mRNA levels in NSCLC and corresponding paratumor tissues. Tissue microarrays (TMA) analysis of 208 NSCLC cases were used to evaluate the relationship between RNF38 expression and clinical implications. Prognostic value was assessed by Kaplan-Meier analysis and log-rank tests. Wound-healing assays, trans-well assays, colony formation assays and CCK8 were used to assess cell migration, invasion and proliferative ability respectively. The analysis of epithelial-to-mesenchymal transition (EMT) phenotype was carried out by immunofluorescence and western blot. Results: Our data revealed that elevated RNF38 expression were more common in NSCLC tissues than paired normal tissues in both mRNA (2.82 ± 0.29 vs. 1.23 ± 0.13) and protein (2.75 ± 0.09 vs. 1.24 ± 0.02) level. High levels of RNF38 expression were significantly associated with lymph node metastases, higher TNM stages (p=0.011), larger tumor size (p=2.09E-04) and predicted poor prognosis. RNF38 expression was inversely correlated with E-cadherin expression (P= 0.025). Moreover, downregulation of RNF38 impaired the proliferation, metastatic and invasive abilities in NSCLC cells. In addition, aberrant RNF38 expression could modulate the key molecules of EMT. Conclusions: Our results indicate that elevated expression of RNF38 is significantly associated with the proliferation and metastatic capacity of NSCLC cells, and RNF38 overexpression can serve as a biomarker of NSCLC poor prognosis.

Adoptive immunotherapy shows encouraging benefit on non-small cell lung cancer: a systematic review and meta-analysis

Although adoptive immunotherapy (AIT) is a novel emerging target treatment for non-small cell lung cancer (NSCLC), its actual efficacy remains controversial. In this meta-analysis, we aimed to evaluate the efficacy of AIT for NSCLC. We systematically searched PubMed, the Cochrane Library, EMBASE, Medline, and Web of Science for relevant parallel randomized controlled trials (RCTs) and high-quality observation studies of AIT without any language restrictions. Two investigators reviewed all the texts and extracted information regarding overall survival rate (OS), progression-free survival rate (PFS), objective response rate (ORR), and disease control rate (DCR) from eligible studies; sensitivity analyses and subgroup analyses were also conducted to reduce heterogeneity Of 319 suitable studies, 15 studies (13 RCTs and 2 observation studies) involving 1684 patients were finally included. Compared to the Control therapy (CT) group, the AIT group exhibited better 1-year OS (P = 0.001), 2-year OS (P < 0.001), 3-year OS (P < 0.001), 5-year OS (P = 0.032), 1-year PFS (P < 0.001), and 2-year PFS (P = 0.029). The difference in the ORR (P = 0.293) and DCR (P = 0.123) was not significant between the groups. The subgroup analysis showed that DC/CIK did more benefit to NSCLC patients than LAK and the cycles not associated with AIT efficacy. AIT can significantly improve the OS and PFS with acceptable toxicity for NSCLC. Nevertheless, further multicenter studies are needed to confirm our conclusion and determine which adoptive immunotherapy is associated with the greatest efficacy.Although adoptive immunotherapy (AIT) is a novel emerging target treatment for non-small cell lung cancer (NSCLC), its actual efficacy remains controversial. In this meta-analysis, we aimed to evaluate the efficacy of AIT for NSCLC. We systematically searched PubMed, the Cochrane Library, EMBASE, Medline, and Web of Science for relevant parallel randomized controlled trials (RCTs) and high-quality observation studies of AIT without any language restrictions. Two investigators reviewed all the texts and extracted information regarding overall survival rate (OS), progression-free survival rate (PFS), objective response rate (ORR), and disease control rate (DCR) from eligible studies; sensitivity analyses and subgroup analyses were also conducted to reduce heterogeneityOf 319 suitable studies, 15 studies (13 RCTs and 2 observation studies) involving 1684 patients were finally included. Compared to the Control therapy (CT) group, the AIT group exhibited better 1-year OS (P = 0.001), 2-year OS (P < 0.001), 3-year OS (P < 0.001), 5-year OS (P = 0.032), 1-year PFS (P < 0.001), and 2-year PFS (P = 0.029). The difference in the ORR (P = 0.293) and DCR (P = 0.123) was not significant between the groups. The subgroup analysis showed that DC/CIK did more benefit to NSCLC patients than LAK and the cycles not associated with AIT efficacy.AIT can significantly improve the OS and PFS with acceptable toxicity for NSCLC. Nevertheless, further multicenter studies are needed to confirm our conclusion and determine which adoptive immunotherapy is associated with the greatest efficacy.