Kilohertz high-frequency electrical stimulation ameliorate hyperalgesia by modulating transient receptor potential vanilloid-1 and N-methyl-D-aspartate receptor-2B signaling pathways in chronic constriction injury of sciatic nerve mice.

The number of patients with neuropathic pain is increasing in recent years, but drug treatments for neuropathic pain have a low success rate and often come with significant side effects. Consequently, the development of innovative therapeutic strategies has become an urgent necessity. Kilohertz High Frequency Electrical Stimulation (KHES) offers pain relief without inducing paresthesia. However, the specific therapeutic effects of KHES on neuropathic pain and its underlying mechanisms remain ambiguous, warranting further investigation. In our previous study, we utilized the Gene Expression Omnibus (GEO) database to identify datasets related to neuropathic pain mice. The majority of the identified pathways were found to be associated with inflammatory responses. From these pathways, we selected the transient receptor potential vanilloid-1 (TRPV1) and N-methyl-D-aspartate receptor-2B (NMDAR2B) pathway for further exploration. Mice were randomly divided into four groups: a Sham group, a Sham/KHES group, a chronic constriction injury of the sciatic nerve (CCI) group, and a CCI/KHES stimulation group. KHES administered 30 min every day for 1 week. We evaluated the paw withdrawal threshold (PWT) and thermal withdrawal latency (TWL). The expression of TRPV1 and NMDAR2B in the spinal cord were analyzed using quantitative reverse-transcriptase polymerase chain reaction, Western blot, and immunofluorescence assay. KHES significantly alleviated the mechanical and thermal allodynia in neuropathic pain mice. KHES effectively suppressed the expression of TRPV1 and NMDAR2B, consequently inhibiting the activation of glial fibrillary acidic protein (GFAP) and ionized calcium binding adapter molecule 1 (IBA1) in the spinal cord. The administration of the TRPV1 pathway activator partially reversed the antinociceptive effects of KHES, while the TRPV1 pathway inhibitor achieved analgesic effects similar to KHES. KHES inhibited the activation of spinal dorsal horn glial cells, especially astrocytes and microglia, by inhibiting the activation of the TRPV1/NMDAR2B signaling pathway, ultimately alleviating neuropathic pain.

Analysis of genotypes and biochemical phenotypes of neonates with abnormal metabolism of butyrylcarnitine. / ä¸é °åŸºè‚‰ç¢±ä»£è°¢å¼‚å¸¸æ–°ç”Ÿå„¿çš„åŸºå› åž‹å’Œç”ŸåŒ–è¡¨åž‹åˆ†æž.

OBJECTIVES: To investigate the genotypes and biochemical phenotypes of neonates with abnormal metabolism of butyrylcarnitine (C4). METHODS: One hundred and twenty neonates with increased C4 levels detected by tandem mass spectrometry in the neonatal screening at Children's Hospital, Zhejiang University School of Medicine from January 2018 to June 2023 were included. The initial screening data and recalled data of C4 and C4/C3 were collected and converted into multiples of C4 reference range. Next generation sequencing was performed and the exons with adjacent 50 bp regions of ACAD8 and ACADS genes were captured by liquid phase capture technique. Variant information was obtained by bioinformatic analysis and the pathogenicity were classified according to the American College of Medical Genetics and Genomics criteria. The Wilcoxon rank sum test was used to analyze the differences in C4 levels among neonates with different variation types. RESULTS: In total, 32 variants in ACAD8 gene were detected, of which 7 variants were reported for the first time; while 41 variants of ACADS gene were detected, of which 17 variants have not been previously reported. There were 39 cases with ACAD8 biallelic variations and 3 cases with ACAD8 monoallelic variations; 34 cases with ACADS biallelic variations and 36 cases with ACADS monoallelic variations. Furthermore, 5 cases were detected with both ACAD8 and ACADS gene variations. Inter group comparison showed that the multiples of C4 reference range in initial screening and re-examination of the ACAD8 biallelic variations and ACADS biallelic variations groups were significantly higher than those of the ACADS monoallelic variations group (all P<0.01), while the multiples in the ACAD8 biallelic variations group were significantly higher than those in the ACADS biallelic variations group (all P<0.01). The multiples of C4 reference range in the initial screening greater than 1.5 times were observed in all neonates carrying ACAD8 or ACADS biallelic variations, while only 25% (9/36) in neonates carrying ACADS monoallelic variations. CONCLUSIONS: ACAD8 and/or ACADS gene variants are the main genetic causes for elevated C4 in newborns in Zhejiang region with high genotypic heterogeneity. The C4 levels of neonates with biallelic variations are significantly higher than those of neonates with monoallelic variations. The cut-off value for C4 level could be modestly elevated, which could reduce the false positive rate in tandem mass spectrometry neonatal screening.

Application of genetic screening processor in screening neonatal glucose-6-phosphate dehydrogenase deficiency.

To evaluate the performance of genetic screening processor (GSP analyzer) in neonatal screening for glucose-6-phosphate dehydrogenase (G6PD)deficiency. The accuracy and precision of GSP analyzer was evaluated with the control materials from National Center for Clinical Laboratories and the low and high quality G6PD control kit (fluorescence analysis). GSP analyzer and semi-automatic fluorescence immunoanalyzer (1420 analyzer) were simultaneously used to detect 2622 neonatal screening samples and 41 confirmed samples to analyze the correlation and consistency of the test results; 78 floating samples and 78 non-floating samples were detected to compare the result. A total of 1 100 384 neonatal screening samples from January 2017 to December 2018 and 855 856 neonatal screening samples from January 2019 to December 2020 were detected with 1420 analyzer and GSP analyzer, respectively. Referring to the percentile method and the expert consensus, the new cut-off value of GSP analyzer for G6PD deficiency in screening was established. The relative bias of GSP analyzer in detecting G6PD was 0.71%-4.23%; the intra assay precision was 4.34%-4.91%, the inter assay precision was 0.85%-2.12%, and the total coefficient of variation was 5.44%-5.72%. There was a significant positive correlation between G6PD activity detected by GSP analyzer and 1420 analyzer (=0.740, <0.01). Forty-one clinical confirmed patients were identified by both 1420 analyzer and GSP analyzer (=0.945). The G6PD activity in floating dry blood spots detected by 1420 analyzer was significantly lower than that in non-floating dry blood spots (<0.05), but there was no significant difference in G6PD activity between floating and non-floating dry blood spots detected by GSP analyzer (>0.05). The sensitivities of GSP analyzer and 1420 analyzer in screening G6PD deficiency were both 100.00%, and the specificities were both more than 99.80%. Compared with 1420 analyzer, the positive predictive value, positive rate and prevalence of G6PD deficiency detected by GSP analyzer were increased, and the false positive rate was decreased (all <0.01). The new cut-off value was 26.1 U/dL for male and 29.1 U/dL for female according to the 99.1% percentile of the population. GSP analyzer has better detection performance with high automation, efficiency and throughput, which can be used in large-scale screening for neonatal G6PD deficiency.

Mechanical and vibrational behaviors of bilayer hexagonal boron nitride in different stacking modes.

Hexagonal boron nitride (h-BN) is a semiconductor material with a wide band gap, which has great potential to serve as a nanoresonators in microelectronics and mass and force sensing fields. This paper investigates the mechanical properties and natural frequencies of bilayer h-BN nanosheets under five different stacking modes, which have been rarely studied, using molecular dynamics simulations. The mechanical properties, including Young's modulus, the ultimate stress, ultimate strain, Poisson's ratio and shear modulus, are studied for all five stacking modes. And the effects of strain rate, crystal orientation and temperature to bilayer h-BN nanosheets' tensile properties have also been studied. Our findings suggest that bilayer h-BN nanosheets are basically an anisotropic material whose tensile properties vary substantially with stacking modes and temperature. Moreover, the natural frequencies are proposed in an explicit form based on the nonlocal theory. The differences of the fundamental natural frequencies among different stacking modes are affected by the constraint condition of bilayer h-BN sheet. The theory results match well with the simulation results. These findings establish elementary understandings of the mechanical behavior and vibration character of bilayer h-BN nanosheets under five different stacking modes, which could benefit its application in advanced nanodevices.

High-frequency electrical stimulation reduced hyperalgesia and the activation of the Myd88 and NFκB pathways in chronic constriction injury of sciatic nerve-induced neuropathic pain mice.

Neuropathic pain has become a global public problem and health burden. Pharmacological interventions are the primary treatment, but the drug cure rate is low with side effects. There is an urgent need to develop novel treatment approaches. High frequency electrical stimulation (KHES) has been widely applied in clinical analgesia. However, its mechanism is poorly understood. In this study, datasets related to neuropathic pain were obtained from the GEO database. The differentially expressed genes (DEGs) and key genes were analyzed through functional enrichment analysis, showing that most of the pathways involve the inflammation. The MyD88 and NFκB pathways were further studied. KHES significantly alleviated mechanical and thermal allodynia in chronic constriction injury of the sciatic nerve mice. KHES also inhibited the increase in Myd88 and p-NFκB expression. The administration of NFκB pathway activator partly reversed the antinociceptive effects of KHES, and NFκB pathway inhibitor achieved analgesic effects similar to those of KHES. Therefore, KHES might be a novel intervention for the treatment of neuropathic pain.

miR-31/QKI-5 axis facilitates cell cycle progression of non-small-cell lung cancer cells by interacting and regulating p21 and CDK4/6 expressions.

BACKGROUND: RNA-binding protein Quaking-5 (QKI-5), a major isoform of QKIs, inhibits tumor progression in non-small cell lung cancer (NSCLC). However, the underlying molecular mechanisms of QKI-5 in the cell cycle of NSCLC are still largely unknown. METHODS: MTT, flow cytometry, and colony formation assays were used to investigate cellular phenotypic changes. Mice xenograft model was used to evaluate the antitumor activities of QKI-5. Co-immunoprecipitation, RNA immunoprecipitation (RIP), and RIP sequencing were used to investigate protein-protein interaction and protein-mRNA interaction. RESULTS: The QKI-5 expression was downregulated in NSCLC tissues compared with that in paired normal adjacent lung tissues. Overexpression of QKI-5 inhibited NSCLC cell proliferative and colony forming ability. In addition, QKI-5 induced cell cycle arrest at G0/G1 phase through upregulating p21Waf1/Cip1 (p21) expression and downregulating cyclin D1, cyclin-dependent kinase 4 (CDK4), and CDK6 expressions. Further analyses showed that QKI-5 interacts with p21 protein and CDK4, CDK6 mRNAs, suggesting a critical function of QKI-5 in cell cycle regulation. In agreement with in vitro study, the mouse xenograft models validated tumor suppressive functions of QKI-5 in vivo through altering cell cycle G1-phase-associated proteins. Moreover, we demonstrated that QKI-5 is a direct target of miR-31. The QKI-5 expression was anticorrelated with the miR-31 expression in NSCLC patient samples. CONCLUSION: Our results suggest that the miR-31/QKI-5/p21-CDK4-CDK6 axis might have critical functions in the progression of NSCLC, and targeting this axis could serve as a potential therapeutic strategy for NSCLC.

Untargeted Metabolomics Analysis Revealed the Difference of Component and Geographical Indication Markers of Panax notoginseng in Different Production Areas.

Panax notoginseng (P. notoginseng) has excellent medicinal and food dual-use characteristics. However, P. notoginseng with a unique origin label has become the target of fraud because of people confusing or hiding its origin. In this study, an untargeted nuclear magnetic resonance (NMR)-based metabolomics approach was used to discriminate the geographical origins of P. notoginseng from four major producing areas in China. Fifty-two components, including various saccharides, amino acids, saponins, organic acids, and alcohols, were identified and quantified through the NMR spectrum, and the area-specific geographical identification components were further screened. P. notoginseng from Yunnan had strong hypoglycemic and cardiovascular protective effects due to its high acetic acid, dopamine, and serine content, while P. notoginseng from Sichuan was more beneficial for diseases of the nervous system because of its high content of fumarate. P. notoginseng from Guizhou and Tibet had high contents of malic acid, notoginsenoside R1, and amino acids. Our results can help to distinguish the geographical origin of P. notoginseng and are readily available for nutritional recommendations in human consumption.

Acid-induced Poria cocos alkali-soluble polysaccharide hydrogel: Gelation behaviour, characteristics, and potential application in drug delivery.

Poria cocos alkali-soluble polysaccharide (PCAP), a water-insoluble ß-glucan, is the main component of the total dried sclerotia of Poria cocos. However, its gelation behaviour and properties have yet to be comprehensively studied. In this study, an acid-induced physical hydrogel based on natural PCAP is fabricated. The acid-induced gelation in PCAP is explored with respect to the pH and polysaccharide concentration. PCAP hydrogels are formed in the pH range of 0.3-10.5, and the lowest gelation concentration is 0.4 wt%. Furthermore, dynamic rheological, fluorescence, and cyclic voltammetry measurements are performed to elucidate the gelation mechanism. The results reveal that hydrogen bonds and hydrophobic interactions play a dominant role in gel formation. Subsequently, the properties of the PCAP hydrogels are investigated using rheological measurements, scanning electron microscopy, gravimetric analysis, free radical scavenging, MTT assays, and enzyme-linked immunosorbent assays. The PCAP hydrogels exhibit a porous network structure and cytocompatibility, in addition to good viscoelastic, thixotropic, water-holding, swelling, antioxidant, and anti-inflammatory activities. Furthermore, using rhein as a model drug for encapsulation, it is demonstrated that its cumulative release behaviour from the PCAP hydrogel is pH dependent. These results indicate the potential of PCAP hydrogels for application in biological medicine and drug delivery.

Multidimensional study of cell division cycle-associated proteins with prognostic value in gastric carcinoma.

Gastric cancer (GC) represents a widespread malignancy with a poor prognosis. Hence, discovering reliable biomarkers is necessary. The cell division cycle-associated protein (CDCA) family, comprising CDCA1-8, plays a key role in tumor progression. However, whether CDCA expression has prognostic value in GC, especially stomach adenocarcinoma (STAD), has not been elucidated yet. Consequently, we conducted a multifaceted study using bioinformatic tools aimed at exploring CDCA expression levels and appraising their potential prognostic values in patients with STAD. All eight CDCAs were significantly upregulated in STAD tissues compared with healthy tissues. Elevated CDCA4/7/8 mRNA expression predicted a short overall survival, and increased CDCA7 transcriptional levels predicted a short disease-free survival. The most frequent alteration in patients with STAD was low mRNA expression. The functional enrichment analysis incorporating the gene ontology (GO) and Kyoto encyclopedia of genes and genomes (KEGG) pathways showed that the cell cycle, foxO signaling pathway, and Epstein-Barr virus were relevant to the main functions of CDCAs. Finally, the immune infiltration analysis revealed a significant correlation between CDCA expression and the infiltration extent of six immunocytes. Therefore, differentially expressed CDCAs may represent potential biomarkers for the prognosis of patients with STAD that can improve survival. Furthermore, this study might offer new ideas for the design and development of immunotherapeutic drugs.

Trends in accessibility of negotiated targeted anti-cancer medicines in Nanjing, China: An interrupted time series analysis.

Background: In order to establish a long-term strategy for bearing the costs of anti-cancer drugs, the state had organized five rounds of national-level pricing negotiations and introduced the National Health Insurance Coverage (NHIC) policy since 2016. In addition, the National Healthcare Security Administration (NHSA) introduced the volume-based purchasing (VBP) pilot program to Nanjing in September 2019. Taking non-small cell lung cancer as an example, the aim of the study was to verify whether national pricing negotiations, the NHIC policy and the VBP pilot program had a positive impact on the accessibility of three targeted anti-cancer drugs. Methods: Based on the hospital procurement data, interrupted time series (ITS) design was used to analyze the effect of the health policy on the accessibility and affordability of gefitinib, bevacizumab and recombinant human endostatin from January 2013 to December 2020 in Nanjing, China. Results: The DDDs of the three drugs increased significantly after the policy implementation (P < 0.001, P < 0.001, P = 0.008). The trend of DDDc showed a significant decrease (P < 0.001, P < 0.001, P < 0.001). The mean availability of these drugs before the national pricing negotiation was <30% in the surveyed hospitals, and increased significantly to 60.33% after 2020 (P < 0.001, P = 0.001, P < 0.001). The affordability of these drugs has also increased every year after the implementation of the insurance coverage policy. The financial burden is higher for the rural patients compared with the urban patients, although the gap is narrowing. Conclusion: The accessibility of targeted anti-cancer drugs has increased significantly after the implementation of centralized prices, the NHIC policy and the VBP pilot program, and has shown sustained long-term growth. Multi-pronged supplementary measures and policy approaches by multiple stakeholders will facilitate equitable access to effective and affordable anti-cancer drugs.

Potential prognostic value of the lymph node ratio and its correlation with circulating sex hormone concentration in pathological T1/2 breast cancer patients: a retrospective study.

Background: The lymph node ratio (LNR) is an additional informative factor complementing anatomic TNM staging in breast cancer patients. The aim of this study was to evaluate the role of LNR in the cancer-specific and overall survival (OS) in a cohort of pT1/2 breast cancer patients and examine its correlation with circulating sex hormone concentrations in postmenopausal cases of the cohort from eastern China islands. Methods: Clinical and pathological characteristics, preoperational sex hormone and tumor markers concentrations, and breast cancer-specific survival (BCSS) and OS were analyzed retrospectively in 732 pathological T1/2 breast cancer patients. Results: The LNR was calculated, and the cut-off value was defined as 0.042 by receiver operative characteristic (ROC) curve according to the patient's mortalities. Patients with LNR ≥0.042 exhibited worse BCSS and OS than others (P<0.001) in pT1/2 breast cancer. Among patients with non-triple negative breast cancer (TNBC) and TNBC subtypes, the LNR ≥0.042 group also exhibited worse BCSS and OS than the LNR <0.042 group (P=0.003, 0.001, and P=0.032, 0.001, respectively). In univariate analysis, unfavorable BCSS and OS were both related with LNR ≥0.042 (P=0.001, <0.001). However multivariate analysis demonstrated TNBC subtypes were independent predictor for BCSS and OS [hazard ratio (HR) =1.449, 95% CI: 1.097-1.914, P=0.009; HR =1.365, 95% CI: 1.093-1.705, P=0.006, respectively]. Notably, Pearson or spearman correlation analysis revealed follicle-stimulating hormone (FSH) and, luteinizing hormone (LH) levels were significantly negatively associated with the LNR (P=0.007, 0.011, respectively) in postmenopausal cases, whereas CA153, CA125 and CEA were positively correlated with it (P<0.001, <0.001, 0.001, respectively) in all cases. Conclusions: Among pT1/2 breast cancer patients from eastern China islands, the LNR is a predictive prognosis factor; a higher LNR seems to correlate with a worse survival outcome both overall and in the subgroups. Strikingly, the current results reveal that serum FSH and LH level inversely associated with axillary node invasion in postmenopausal cases, whereas tumor markers directly related with it. The LNR is an informative factor complementing TNM staging.

Prevalence of Feline Immunodeficiency Virus and Toxoplasma gondii in Feral Cats on St. Kitts, West Indies.

Toxoplasma gondii (T. gondii) is a cosmopolitan protozoan parasite that infects all warm-blooded species including humans. The definitive hosts of T. gondii are felid vertebrates including the domestic cat. Domestic cats shed oocysts for approximately two weeks in their feces after the primary infection. It has been shown that feline immunodeficiency virus (FIV) positive cats have a higher prevalence of and a higher titer of antibodies to T. gondii than those of FIV-negative cats. The main purposes of this study were to determine FIV prevalence and to investigate the oocysts shedding in FIV-positive and FIV-negative feral cats on St. Kitts. Fecal samples were collected from feral cats while their FIV statues were determined using a commercial SNAP kit. Total fecal DNA of each cat was tested for the presence of T. gondii DNA using a polymerase chain reaction (PCR) consistently detecting one genome equivalent. A FIV-positive status was detected in 18 of 105 (17.1%, 95% confidence interval (CI): 9.9%-24.3%) feral cats sampled. Furthermore, males were three times more likely to be FIV positive than females (p = 0.017) with an odds ratio of 3.93 (95% CI: 1.20-12.89). Adults were found to have at least twice the prevalence of FIV compared to cats younger than one year of age (p = 0.056) with an odds ratio of 3.07 (95% CI: 0.94-10.00). Toxoplasma gondii DNA was not detected in the feces of any of the 18 FIV-positive (95% CI: 0%-0.18%) and 87 FIV-negative cats (95% CI: 0%-0.04%). A follow-up study with a much bigger sample size is needed to prove or disprove the hypothesis that FIV-positive cats have a higher prevalence of shedding T. gondii oocysts than FIV-negative cats.

Small Indian Mongooses (Herpestes auropunctatus) Serve As Reservoirs of Bartonella henselae and Rickettsia felis Vectored by Ctenocephalides felis.

Small Indian mongooses (SIMs, Herpestes auropunctatus) have invasively inhabited over 60 islands worldwide. They have been confirmed as a reservoir of rabies, leptospirosis, and salmonellosis; however, their role in the epidemiology of other zoonoses is little known. On St. Kitts, as well as other islands, SIMs harbor Ctenocephalides felis, which can vector several zoonotic diseases. In this study, SIMs were examined for fleas, and the collected fleas analyzed by PCR and DNA sequencing for Bartonella henselae, Rickettsia felis, Yersinia pestis, and Dipylidium caninum. Of the 87 SIMs, 75 (86.2%) harbored C. felis. C. felis recovered from nine (10.3%), one (1.1%), and one (1.1%) of the SIMs was positive for B. henselae, R. felis, and D. caninum, respectively. These data indicate that SIMs serve as an additional reservoir of B. henselae and R. felis, which should be taken into consideration in control and prevention of these rapidly emerging zoonoses.

Effectively Intervening Epithelial-Mesenchymal Transition of Retinal Pigment Epithelial Cells With a Combination of ROCK and TGF-ß Signaling Inhibitors.

Purpose: Epithelial-mesenchymal transition (EMT) of retinal pigment epithelial (RPE) cells is a key pathological event in proliferative retinal diseases such as proliferative vitreoretinopathy (PVR). This study aimed to explore a new method to reverse EMT in RPE cells to develop an improved therapy for proliferative retinal diseases. Methods: In vitro, human embryonic stem cell-derived RPE cells were passaged and cultured at low density for an extended period of time to establish an EMT model. At different stages of EMT after treatment with known molecules or combinations of molecules, the morphology was examined, transepithelial electrical resistance (TER) was measured, and expression of RPE- and EMT-related genes were examined with RT-PCR, Western blotting, and immunofluorescence. In vivo, a rat model of EMT in RPE cells was established via subretinal injection of dispase. Retinal function was examined by electroretinography (ERG), and retinal morphology was examined. Results: EMT of RPE cells was effectively induced by prolonged low-density culture. After EMT occurred, only the combination of the Rho-associated coiled-coil containing protein kinase (ROCK) inhibitor Y27632 and the TGF-ß receptor inhibitor RepSox (RY treatment) effectively suppressed and reversed the EMT process, even in cells in an intermediate state of EMT. In dispase-treated Sprague-Dawley rats, RY treatment maintained the morphology of RPE cells and the retina and preserved retinal function. Conclusions: RY treatment might promote mesenchymal-epithelial transition (MET), the inverse process of EMT, to maintain the epithelial-like morphology and function of RPE cells. This combined RY therapy could be a new strategy for treating proliferative retinal diseases, especially those involving EMT of RPE cells.

Kilohertz high-frequency alternating current blocks nerve conduction without causing nerve damage in rats.

BACKGROUND: In recent years, 2-50 kHz high-frequency alternating current has been shown to block nerve conduction mostly based on simulation models or experiments in vitro. This study aimed to assess the nerve block effects and related parameters of kilohertz alternating current in a rat model. METHODS: High-frequency biphasic rectangular stimulus pulse was applied to rat's sciatic nerve in vivo, and its blockade frequency and intensity was studied by recording the changes of compound muscle action potential (CMAP) amplitude and muscle states before and after stimulation. Secondly, diameter and circumference of sciatic nerve was measured at stimulating point by ultrasound. The correlation between stimulus' frequency and the nerve's diameter and circumference was studied. Lastly, we assessed nerve damage causing by high-frequency electrical stimulation by measuring CMAP and nerve conduction velocity (NCV) in the following day and sciatic nerve hematoxylin-eosin staining, both blocked side and contralateral side. RESULTS: When the current intensity was fixed, the blockade only occurred in a specific frequency range, above or below might have partial block effect. Preliminary statistical results showed that the blocking frequency of high-frequency alternating current was negatively linearly correlated with the circumference of sciatic nerve (P<0.05); HE staining of the sciatic nerve showed no axon and myelin sheath damage on blocked or opposite side, and the CMAP and NCV of the sciatic nerve remeasured in the next day were normal, indicating high-frequency electrical stimulation produced no nerve injury. CONCLUSIONS: High-frequency alternating current stimulation can block nerve conduction without causing nerve damage, and the complete block frequency is negatively linearly correlated with the circumference of sciatic nerve.

A Prediction Rule for Overall Survival in Non-Small-Cell Lung Cancer Patients with a Pathological Tumor Size Less Than 30 mm.

We sought to develop and validate a clinical nomogram model for predicting overall survival (OS) in non-small-cell lung cancer (NSCLC) patients with resected tumors that were 30 mm or smaller, using clinical data and molecular marker findings. We retrospectively analyzed 786 NSCLC patients with a pathological tumor size less than 30 mm who underwent surgery between 2007 and 2017 at our institution. We identified and integrated significant prognostic factors to build the nomogram model using the training set, which was subjected to the internal data validation. The prognostic performance was calibrated and evaluated by the concordance index (C-index) and risk group stratification. Multivariable analysis identified the pathological tumor size, lymph node metastasis, and Ki-67 expression as independent prognostic factors, which were entered into the nomogram model. The nomogram-predicted probabilities of OS at 1 year, 3 years, and 5 years posttreatment represented optimal concordance with the actual observations. Harrell's C-index of the constructed nomogram with the training set was 0.856 (95% CI: 0.804-0.908), whereas TNM staging was 0.814 (95% CI: 0.742-0.886, P = 5.280221e - 13). Survival analysis demonstrated that NSCLC subgroups showed significant differences in the training and validation sets (P < 0.001). A nomogram model was established for predicting survival in NSCLC patients with a pathological tumor size less than 30 mm, which would be further validated using demographic and clinicopathological data. In the future, this prognostic model may assist clinicians during treatment planning and clinical studies.

Prognostic value of mutant p53, Ki-67, and TTF-1 and their correlation with EGFR mutation in patients with non-small cell lung cancer.

INTRODUCTION: The clinical characteristics of non-small cell lung cancer (NSCLC) with epidermal growth factor receptor (EGFR) mutation have been well studied. However, the correlation of EGFR mutation with mutant p53, Ki-67, and thyroid transcription factor 1 (TTF-1) and their prognostic value remain indistinct. MATERIAL AND METHODS: Clinical and pathological characteristics and overall survival were analysed retrospectively in 523 surgically resected NSCLC patients. The expression levels of p53, Ki-67, and TTF-1 protein were detected by immunohistochemistry, and an amplification refractory mutation system was used to access the status of EGFR mutations. RESULTS: Of 523 patients with surgically resected NSCLC, 210 patients (38.4%) harboured EGFR mutations. Compared to the EGFR wild-type lung cancer, mutated EGFR harboured significantly increased mutant p53-positive or TTF-1-positive tumors (P<0.001 and<0.001, respectively). Former or current smokers, pathological stage and mutant p53-or TTF-1-positive status were independent predictors of EGFR mutation (P=0.001, 0.014, 0.014 and <0.001, respectively). Patients with p53 under expression had significantly better overall survival in the whole cohort and wild-type EGFR cohort (P=0.0010 and 0.0020, respectively) as well as in Ki-67-negative and TTF-1-positive patients (P<0.0001 and 0.0009, and P<0.0001 and 0.0004, respectively). Interestingly, in patients harbouring EGFR mutations, p53-under expression and Ki-67-negative cases still had better survival than positive cases, whereas there was no obvious difference between TTF-1-negative and TTF-1-positive cases (P=0.0198, 0.0068 and 0.3684, respectively). Finally, in NSCLC patients with wild-type EGFR, positive Ki-67 expression was the independent predictor for the worst survival (P=0.022). CONCLUSION: The expression levels of mutant p53, Ki-67, and TTF-1 were correlated with EGFR mutation. High expression of mutant p53 and Ki-67 correlated with poor survival in the entire cohort, EGFR mutation or wild-type cohort. In addition, Ki-67 might have an impact on the prognosis for patients with NSCLC with wild-type EGFR.

Epidermal Growth Factor Receptor Mutations and Their Prognostic Value with Carcinoembryonic Antigen in Pathological T1 Lung Adenocarcinoma.

AIMS: The prognostic value of epidermal growth factor receptor (EGFR) mutations in the context of serum carcinoembryonic antigen levels remains controversial in T1 lung adenocarcinoma. METHODS: Clinical and pathological characteristics, preoperational carcinoembryonic antigen levels, EGFR mutations, and disease-free and overall survival were analysed retrospectively in 573 pathological T1 patients in East China. RESULTS: EGFR mutations were detected in 220 of 573 patients (38.4%). Patients with serum carcinoembryonic antigen levels ≥ 2.12 ng/mL had worse disease-free (P < 0.001) and overall survival (P < 0.001) than had others, although survival was comparable between patients with and without EGFR mutations. However, patients with exon 21 mutations in EGFR had significantly better overall survival than had patients with exon 19 mutations (P = 0.016), although disease-free survival was comparable (P = 0.424). Among patients with serum carcinoembryonic antigen levels ≥ 2.12 ng/mL, disease-free (P = 0.019) and overall survival (P < 0.001) was also better than that in those with exon 21 mutations. Finally, the exon 19 deletion was found to be an independent predictor of unfavourable overall survival (P = 0.037). CONCLUSIONS: EGFR mutations were associated with preoperational serum carcinoembryonic antigen levels ≥ 2.12 ng/mL. In patients with levels above this threshold, those with the exon 19 deletion have less favourable prognosis than have those with the exon 21 mutation.

Depression Screening in an Academic Family Practice.

BACKGROUND AND OBJECTIVES: Screening for depression in primary care can be effective, but ensuring that appropriate care is available and engaging patients in treatment are major challenges. Even when follow-up care is available, patient engagement often relies on the primary care provider initiating care. In this study we wanted to assess the effectiveness of a depression screening program in an academic family practice. RESULTS: Depression screening occurred in 98.4% of all adult encounters (n=3,341). Of these patients, 7.3% screened positive for depression and were not presenting for mood problems. Only 33.7% of patients with positive screens had their results addressed. Patients who had their results addressed were twice as likely to return for follow-up as those who did not (34.1% versus 17.4%). Patients with severe depression were more likely to follow-up than patients with mild depression (53% versus 15%). RESULTS: Depression screening occurred in 98.4% of all adult encounters (n=3341). Of these patients 7.3% screened positive for depression and were not presenting for mood problems. Only 33.7% of patients with positive screens had their results addressed. Patients who had their results addressed were twice as likely to return for follow-up as those who did not (34.1% vs. 17.4%, P<.01). Patients with severe depression were more likely to follow-up than patients with mild depression (53% vs. 15%, P<.01). CONCLUSIONS: Depression screening can be efficiently incorporated into primary care practice, but engaging providers and patients in diagnosis and treatment is challenging. We recommend a systems-based approach that emphasizes immediate access to treatment when implementing depression screening in a primary care practice.

Increased expression of T cell immunoglobulin and mucin domain 4 is positively associated with the disease severity of patients with ankylosing spondylitis.

Monocytes and associated cytokines have been shown to be involved in the pathogenesis of ankylosing spondylitis (AS). T cell immunoglobulin and mucin domain 4 (Tim-4) was identified on monocytes/macrophages and dentritic cells (DCs) and plays important roles in regulating the activities of macrophages and DCs. The current study investigated the association between Tim-4 expression and AS. Our results showed that Tim-4 expression on monocytes and Tim-4 level in plasma were highly increased in AS patients than in controls. Furthermore, TNF-α production and bath ankylosing spondylitis disease activity index (BASDAI) have positive relationships with Tim-4 expression in AS patients. High expression of Tim-4 was thought to contribute to the pathogenesis and an underlying mechanism of AS.