MPZL1 suppresses the cancer stem-like properties of lung cancer through ß-catenin/TCF4 signaling.

This study was designed to explore the influence of myelin protein zero-like protein 1 (MPZL1) on the stem-like properties of cancer cells and the underlying mechanism in lung adenocarcinoma. Real-time quantitative polymerase chain reaction (RT-qPCR) was utilized to evaluate mRNA expression level. CCK8, wound healing, and transwell assays were applied to assess cell proliferation, migration, and invasion. Tumorsphere-formation assay was utilized to assess cancer stem cell-like properties. LF3 was used to block the ß-catenin/Transcription factor 4 (TCF-4) signaling. Xenograft nude mouse model was conducted; tumor weight and volume were recorded. Western blot assay was utilized to detect the expression levels of CD44, CD133, ß-catenin, TCF-4, and MPZL1. Following MPZL1 knockdown, the mRNA expression levels of MPZL1, ß-catenin, and TCF-4 were inhibited, while the mRNA expression levels of the above genes were increased after the MPZL1 overexpression. MPZL1 knockdown suppressed cell proliferation, migration, and invasion, reduced the tumorsphere-formation capacity, and restrained the expression levels of CD44 and CD133. However, MPZL1 overexpression promoted the cell proliferation, migration, and invasion, enhanced the tumorsphere-formation capacity, and increased the expression levels of CD44 and CD133. Interestingly, LF3 treatment partially revised the effect of MPZL1 overexpression. These findings were further corroborated by in vivo experiments. We concluded that MPZL1 could suppress the lung adenocarcinoma cells' proliferation, migration, invasion, and lung cancer stem cells characteristics. The underlying mechanism is involved in the activation of ß-catenin/TCF-4 signaling.

WISP3 suppresses ESCC progression by inhibiting the IGF-2-IGF1R-AKT signaling cascade.

Esophageal squamous cell carcinoma (ESCC) is a major health problem worldwide, especially in the Chinese population. However, the intrinsic molecular mechanisms of ESCC progression are largely unclear, thus there is an unmet need to identify essential genes governing this disease. Here, we discovered WISP3, an important member of the CCN family, is markedly downregulated in ESCC tissues compared to the normal esophageal epithelium. Downregulation of WISP3 in cancer tissue correlates with worse overall survival of ESCC patients. Using ESCC cell lines as models, we found that forced expression of WISP3 not only suppressed proliferation and migration of cancer cells in vitro, but also inhibited ESCC tumor growth and metastasis in vivo. On the contrary, WISP3 depletion strongly promoted the tumorigenicity of ESCC cells. Mechanistically, we found that WISP3 negates the activity of AKT via inhibiting the IGF-2-IGF1R signaling cascade, which mediates the tumor-suppressive function of WISP3 in esophageal cancers. Together, we identified a novel factor driving the development of ESCC, and revealed a potential therapeutic target for ESCC treatment.

LEF1-AS1 contributes to proliferation and invasion through regulating miR-544a/ FOXP1 axis in lung cancer.

Long non-coding RNAs (lncRNAs) are increasingly recognized as important regulators in tumor development. This study aims to investigate the potential role oflncRNALEF1-AS1, in the progression of lung cancer. Quantitative real-time PCR (qRT-PCR) and western blot assays showed that LEF1-AS1 was upregulated while miR-544a was downregulated in lung cancer specimens and cells. Overexpression of LEF1-AS1 led to the enhancement of cell proliferation and invasion, revealed by CCK-8 assay and transwell assay. A negative correlation was found between LEF1-AS1 and miR-544a. BLAST analysis and dual-luciferase assay confirmed that FOXP1 is a downstream effector of miR-544a. Therefore, the LEF1-AS1/miR-544a/FOXP1 axis is an important contributor to lung cancer progression. Collectively, our novel data uncovers a new mechanism that governs tumor progression in lung cancer and provides new targets that may be used for disease monitoring and therapeutic intervention of lung cancer.

Surface reconstruction of pyrite-type transition metal sulfides during oxygen evolution reaction.

Reconstruction universally occurs over non-layered transition metal sulfides (TMSs) during oxygen evolution reaction (OER), leading to the formation of active species metal (oxy)hydroxide and thus significantly influences the OER performance. However, the reconstruction process and underlying mechanism quantitatively remain largely unexplored. Herein, we proposed an electrochemical reaction mechanism, namely sulfide oxidation reaction (SOR), to elucidate the reconstruction process of pyrite-type TMSs. Based on this mechanism, we evaluated the reconstruction capability of NiS2 doped with transition metals V, Cr, Mn, Fe, Co, Cu, Mo, Ru, Rh, and Ir within different doped systems. Two key descriptors were thus proposed to describe the reconstruction abilities of TMSs: USOR (the theoretical electric potential of SOR) and ΔU (the difference between the theoretical electric potential of SOR and OER), representing the initiation electric potential of reconstruction and the intrinsic reconstruction abilities of TMSs, respectively. Our finding shows that a lower USOR readily initiate reconstruction at a lower potential and a larger ΔU indicating a poorer reconstruction ability of the catalyst during OER. Furthermore, Fe-doped CoS2 was used to validate the rationality of our proposed descriptors, being consistent with the experiment findings. Our work provides a new perspective on understanding the reconstruction mechanism and quantifying the reconstruction of TMSs.

Mn-Based Mullites for Environmental and Energy Applications.

Mn-based mullite oxides AMn2O5 (A = lanthanide, Y, Bi) is a novel type of ternary catalyst in terms of their electronic and geometric structures. The coexistence of pyramid Mn3+-O and octahedral Mn4+-O makes the d-orbital selectively active toward various catalytic reactions. The alternative edge- and corner-sharing stacking configuration constructs the confined active sites and abundant active oxygen species. As a result, they tend to show superior catalytic behaviors and thus gain great attention in environmental treatment and energy conversion and storage. In environmental applications, Mn-based mullites have been demonstrated to be highly active toward low-temperature oxidization of CO, NO, volatile organic compounds (VOCs), etc. Recent research further shows that mullites decompose O3 and ozonize VOCs from -20 °C to room temperature. Moreover, mullites enhance oxygen reduction reactions (ORR) and sulfur reduction reactions (SRR), critical kinetic steps in air-battery and Li-S batteries, respectively. Their distinctive structures also facilitate applications in gas-sensitive sensing, ionic conduction, high mobility dielectrics, oxygen storage, piezoelectricity, dehydration, H2O2 decomposition, and beyond. A comprehensive review from basic physicochemical properties to application certainly not only gains a full picture of mullite oxides but also provides new insights into designing heterogeneous catalysts.

Clinical application of real-time continuous glucose monitoring system during postoperative enteral nutrition therapy in esophageal cancer patients.

BACKGROUND: Enteral nutrition (EN) support therapy increases the risk of abnormal blood glucose (BG). The aim of this study is to evaluate the clinical value of a real-time continuous glucose monitoring (rt-CGM) system in BG monitoring during postoperative EN support therapy in patients with esophageal cancer. METHODS: Patients without diabetes mellitus (DM) with esophageal cancer who planned to receive postoperative EN were enrolled. With the self-monitoring of BG value as the reference BG, the accuracy of rt-CGM was evaluated by the mean absolute relative difference (MARD) value, correlation efficient, agreement analysis, and Parkes and Clarke error grid plot. Finally, paired t tests were used to compare the differences in glucose fluctuations between EN and non-EN days and slow and fast days. RESULTS: The total MARD value of the rt-CGM system was 13.53%. There was a high correlation between interstitial glucose and fingertip capillary BG (consistency correlation efficient = 0.884 [95% confidence interval, 0.874-0.894]). Results of 15/15%, 20/20%, 30/30% agreement analysis were 58.51%, 84.71%, and 99.65%, respectively. The Parkes and Clarke error grid showed that the proportion of the A and B regions were 100% and 99.94%, respectively. The glucose fluctuations on EN days vs non-EN days and on fast days vs slow days were large, and the difference was statistically significant (P < 0.001). CONCLUSION: The rt-CGM system achieved clinical accuracy and can be used as a new option for glucose monitoring during postoperative EN therapy. The magnitude of glucose fluctuation during EN therapy remains large, even in the postoperative population without DM.

Preoperative levels of folate receptor-positive circulating tumor cells in different subtypes of early-stage lung adenocarcinoma: Predictive value for determining extent of surgical resection.

Background: The objective was to measure the correlations of preoperative levels of folate receptor-positive circulating tumor cells (FR+CTCs) with clinical characteristics and histologic subtype in early-stage lung adenocarcinoma, and to determine the predictive value of FR+CTC level in preoperative determination of the extent of surgical resection. Patients and methods: In this retrospective, single-institution, observational study, preoperative FR+CTC levels were measured via ligand-targeted enzyme-linked polymerization in patients with early-stage lung adenocarcinoma. Receiver operating characteristic (ROC) analysis was used to identify the optimal cutoff value of FR+CTC level for prediction of various clinical characteristics and histologic subtypes. Results: No significant difference in FR+CTC level was observed among patients with adenocarcinoma in situ (AIS), minimally invasive adenocarcinoma (MIA), and invasive adenocarcinoma (IAC) (P = 0.813). Within the non-mucinous adenocarcinoma group, no difference was observed among patients with tumors whose predominant growth patterns were lepidic, acinar, papillary, micropapillary, solid, and complex gland (P = 0.053). However, significant differences in FR+CTC level were observed between patients with and without the micropapillary subtype [11.21 (8.22-13.61) vs. 9.85 (7.43-12.63), P = 0.017], between those with and without the solid subtype [12.16 (8.27-14.90) vs. 9.87 (7.50-12.49), P = 0.022], and between those with any of the advanced subtypes (micropapillary, solid, or complex glands) vs. none of these [10.48 (7.83-13.67) vs. 9.76 (7.42-12.42), P = 0.032]. FR+CTC level was also correlated with degree of differentiation of lung adenocarcinoma (P = 0.033), presence of visceral pleural invasion (VPI) of lung carcinoma (P = 0.003), and lymph node metastasis of lung carcinoma (P = 0.035). Conclusion: FR+CTC level is of potential predictive value in determining the presence of aggressive histologic patterns (micropapillary, solid, and advanced subtypes), degree of differentiation, and occurrence of VPI and lymph node metastasis in IAC. Measurement of FR+CTC level combined with intraoperative frozen sections may represent a more effective method of guiding resection strategy in cases of cT1N0M0 IAC with high-risk factors.

MPZL1 Promotes Lung Adenocarcinoma Progression by Enhancing Tumor Proliferation, Invasion, Migration, and Suppressing Immune Function via Transforming Growth Factor-ß1.

Lung cancer (LC) is the leading cause of death worldwide, and lung adenocarcinoma (LUAD) is the most common form of LC. The abnormally high expression of myelin protein zero-like 1 (MPZL1) promotes the malignant progression of various tumors. However, there is no relevant report on the functional role of MPZL1 in LUAU. In this study, we applied Illumina sequencing to screen differentially expressed genes. Subsequently, MPZL1 was selected as hub gene for quantitative real-time polymerase chain reaction (qRT-PCR) and CCK8 assay. The expression level of MPZL1 was analyzed by immunohistochemistry, immunofluorescence, western blot, and qRT-PCR. After silencing or overexpressing MPZL1, CCK8, EDU, clone formation, scratch healing, invasion, and nude mouse tumor-bearing experiments were performed to detect the abilities of cell proliferation, migration, invasion, and tumorigenicity. Moreover, qRT-PCR, western blot, coimmunoprecipitation, and scratch healing assays were conducted to explore the transcriptional regulatory factors of MPZL1. Finally, the relationship between MPZL1 and immunotherapy was explored through public databases and validated in vivo. The results show that a total of 196 high-expressed genes and 496 low-expressed genes were screened. Differential genes are mainly enriched in cell proliferation and division, protein binding, and other pathways and functions. MPZL1 was selected as the hub gene and upregulated in LUAD tissues and cells. Silencing MPZL1 inhibited the cell proliferation and cloning formation, and the growth of tumor. Conversely, overexpression of MPZL1 has the opposite effect. In addition, MPZL1 combines with the transforming growth factor-ß1 to promote the progress of LUAD. Finally, we found that high expression of MPZL1 is negatively correlated with infiltration of CD8+ cells and may lead to immunotherapy resistance. In summary, this study revealed a new mechanism by which MPZL1 promotes LUAD progression by enhancing tumor proliferation, invasion, migration, and suppressing immune function.

Design Rules of a Sulfur Redox Electrocatalyst for Lithium-Sulfur Batteries.

Seeking an electrochemical catalyst to accelerate the liquid-to-solid conversion of soluble lithium polysulfides to insoluble products is crucial to inhibit the shuttle effect in lithium-sulfur (Li-S) batteries and thus increase their practical energy density. Mn-based mullite (SmMn2 O5 ) is used as a model catalyst for the sulfur redox reaction to show how the design rules involving lattice matching and 3d-orbital selection improve catalyst performance. Theoretical simulation shows that the positions of Mn and O active sites on the (001) surface are a good match with those of Li and S atoms in polysulfides, resulting in their tight anchoring to each other. Fundamentally, dz2 and dx2 -y2 around the Fermi level are found to be crucial for strongly coupling with the p-orbitals of the polysulfides and thus decreasing the redox overpotential. Following the theoretical calculation, SmMn2 O5 catalyst is synthesized and used as an interlayer in a Li-S battery. The resulted battery has a high cycling stability over 1500 cycles at 0.5 C and more promisingly a high areal capacity of 7.5 mAh cm-2 is achieved with a sulfur loading of ≈5.6 mg cm-2 under the condition of a low electrolyte/sulfur (E/S) value ≈4.6 µL mg-1 .

Fast-Charging Zn-Air Batteries with Long Lifetime Enabled by Reconstructed Amorphous Multi-Metallic Sulfide.

Developing fast-charging Zn-air batteries is crucial for widening their application but remains challenging owing to the limitation of sluggish oxygen evolution reaction (OER) kinetics and insufficient active sites of electrocatalysts. To solve this issue, a reconstructed amorphous FeCoNiSx electrocatalyst with high density of efficient active sites, yielding low OER overpotentials of 202, 255, and 323 mV at 10, 100, and 500 mA cm-2 , respectively, is developed for fast-charging Zn-air batteries with low charging voltages at 100-400 mA cm-2 . Furthermore, the fabricated 3241.8 mAh (20 mA cm-2 , 25 °C) quasi-solid Zn-air battery shows long lifetime of 500 h at -10 and 25 °C as well as 150 h at 40 °C under charging 100 mA cm-2 . The detailed characterizations combine with density functional theory calculations indicate that the defect-rich crystalline/amorphous ternary metal (oxy)hydroxide forms by the reconstruction of amorphous multi-metallic sulfide, where the electron coupling effect among multi-active sites and migration of intermediate O* from Ni site to the Fe site breaks the scaling relationship to lead to a low theoretical OER overpotential of 170 mV, accounting for the outstanding fast-charging property. This work not only provides insights into designing advanced OER catalysts by the self-reconstruction of the pre-catalyst but also pioneers a pathway for practical fast-charging Zn-air batteries.

[Establishment of a cyanotic congenital heart defect porcine model with decreased pulmonary blood flow].

OBJECTIVE: To establish a porcine model of congenital heart disease with decreased pulmonary blood to explore the morphological changes of immature pulmonary vascular vessels. METHODS: Twenty piglets (one to two-month-old) were randomly divided into three groups: sham-operated group (group S, n = 6), small incisions on the right chest, produced a transient reduction in pulmonary blood; Operation group 1(group T(1), n = 7), small incisions on the right chest, producing artificial atrial defect with self-made dilator and simultaneous banding pulmonary artery to generate a systolic pressure gradient between 20 - 30 mm Hg (1 m Hg = 0.133 kPa); Operation group 2(group T(2), n = 7): operation procedure was similar as group T(1) with systolic pressure gradient between 30 - 50 mm Hg. Lung tissue from right middle lobe (1.0 cm×0.8 cm×0.8 cm) was taken immediately after thoracotomy, at the end of surgery and at 2 months after operation and stained by Weigert (elastic fiber) and van Gieson (collagen) methods to observe the morphological changes. RESULTS: Five animals survived in Group S, 6 animals survived in group T(1) and 5 animals survived in group T(2). The inside diameter of pulmonary arterioles after thoracotomy and at the end of surgery was similar among the three groups (P > 0.05). At 2 months after operation, the inside diameter of pulmonary artery was significantly higher in group T(1) and T(2) than in group S (all P < 0.05) while the number of pulmonary small artery per square centimeter (APSC) of group T(1) and T(2) was significantly lower than that of group S (all P < 0.05). Tunica media of pulmonary artery was thinner and vascular lumen was larger in group T(1) and T(2) compared to those of group S. CONCLUSION: In this piglets model with reduced pulmonary blood, the pulmonary arterioles underwent dysplastic changes. Thus, pulmonary blood flow is an important determinant for the physiological development of pulmonary artery.

Acetylation may strengthen the antitumor activity of low molecular heparin.

BACKGROUND: To synthesize acetylated low anticoagulant low molecular weight heparin (ALMWH) and to detect its antineoplastic activity. METHODS: We obtained Low anticoagulant low molecular weight heparin (LMWH) by splitting unfractionated heparin (UFH) with sodium periodate oxidation and sodium borohydride reduction, then the LMWH was subjected to acetylate catalyzed by dicyclohexylcarbodiimide and dimethylaminopyridine to produce ALMWH. The anti-proliferative activities were determined on MDA-MB-231 human breast cancer cells in vitro. RESULTS: ALMWH exhibited stranger anti-proliferative activity Compared with LMWH, In the MDA-MB-231 cell line, the growth of MDA-MB-231 cells with IC50 of 22.16 µM at 48 h in a concentration-dependent and time-dependent manner, ALMWH produced stronger inhibitory effects especially when it was used in low concentrations. By the use of bulky catalysts, the acetylation site in the molecular chain of low molecular weight heparin with a high selectivity, the synthesis process of Low anticoagulant low molecular weight heparin can be easily controlled. Therefore, large scale industrial production can be carried out. CONCLUSIONS: The synthesized ALMWH possesses a high anti-proliferative activity, Chemical modification of structure can endow LMWH with a high antiproliferative activities. ALMWH is expected to enter clinical trials due to its high druggability. Simultaneously, this study provides a basic method for screening of antineoplastic drug with low toxicity.

Cooperative Catalysis toward Oxygen Reduction Reaction under Dual Coordination Environments on Intrinsic AMnO3 -Type Perovskites via Regulating Stacking Configurations of Coordination Units.

It remains challenging for pure-phase catalysts to achieve high performance during the electrochemical oxygen reduction reaction to overcome the sluggish kinetics without the assistance of extrinsic conditions. Herein, a series of pristine perovskites, i.e., AMnO3 (A = Ca, Sr, and Ba), are proposed with various octahedron stacking configurations to demonstrate the cooperative catalysis over SrMnO3 jointly explored by experiments and first-principles calculations. Comparing with the unitary stacking of coordination units in CaMnO3 or BaMnO3 , the intrinsic SrMnO3 with a mixture of corner-sharing and face-sharing octahedron stacking configurations demonstrates superior activity (Ehalf-wave  = 0.81 V), and charge-discharge stability over 400 h without the voltage gap (≈0.8 V) increasing in zinc-air batteries. The theoretical study reveals that, on the SrMnO3 (110) surface, the active sites switch from coordinatively unsaturated atop Mn (*OO, *OOH) to Mn-Mn bridge (*O, *OH). Therefore, the intrinsic dual coordination environments of Mn-Ocorner and Mn-Oface enable cooperative modulation of the interaction strength of the oxygen intermediates with the surface, inducing the decrease of the *OH desorption energy (rate-limiting step) unrestricted by scaling relationships with the overpotential of ≈0.28 V. This finding provides insights into catalyst design through screening intrinsic structures with multiple coordination unit stacking configurations.

Complete response to crizotinib in a metastatic adenocarcinoma of unknown primary harboring MET amplification and NTRK1 co-occurring mutation.

Carcinomas of unknown primary (CUPs) have poor prognosis due to the paucity of data on their clinical characteristics and laboratory features, and empirical chemotherapy still remains the critical management for this kind of disease. This study aimed to present the knowledge of treating an elderly man with metastatic adenocarcinoma of unknown primary and also with a history of long-term hypertension and renal cysts. He was identified to harbor mesenchymal-epithelial transition factor (MET) gene amplification and neurotrophic tyrosine receptor kinase 1 (NTRK1) gene co-occurring mutation by targeted next-generation sequencing analysis upon the progression of empirical chemotherapy. He was then treated with a standard dose of crizotinib (250 mg, twice daily), which exhibited a satisfactory complete response (CR) of the targeted lesions after 1 month of treatment. When the number of renal cysts increased and renal inadequacy occurred after treatment for 2 months, crizotinib was reduced to half-dose (250 mg, once daily), and still conferred maintenance of CR for another 6.5 months and good quality life of the patient. These results suggested that treatments based on driver genes rather than primary tumor types could be a promising manipulation for achieving better treatment outcome, and a half-dose of crizotinib might be both effective and tolerable for MET-overexpressed CUPs with underlying renal diseases.

Knockdown of TBRG4 affects tumorigenesis in human H1299 lung cancer cells by regulating DDIT3, CAV1 and RRM2.

The transforming growth factor ß regulator 4 (TBRG4) gene, located on the 7p14-p13 chromosomal region, is implicated in numerous types of cancer. However, the contribution(s) of TBRG4 in human lung cancer remains unknown. In the present study, the expression of TBRG4 mRNA was investigated in the H1299 lung cancer cell line using the quantitative polymerase chain reaction (qPCR) following the knockdown of TBRG4 by a lentivirus-mediated small interfering RNA (siRNA). Results identified that the expression of TBRG4 within H1299 cells was significantly suppressed (P<0.01) by RNA interference, and 586 genes were differentially expressed following TBRG4 silencing. Ingenuity Pathway Analysis (IPA) revealed that these genes were often associated with infectious diseases, organismal injury, abnormalities and cancer functional networks. Further IPA of these networks revealed that TBRG4 knockdown in H1299 cells deregulated the expression of 21 downstream genes, including the upregulation of DNA damage-inducible transcript 3 (DDIT3), also termed CCAAT/enhancer-binding protein homologous protein, and downregulation of caveolin 1 (CAV1) and ribonucleotide reductase regulatory subunit M2 (RRM2). Results were validated using qPCR and western blotting. Furthermore, immunohistochemical staining of TBRG4 protein identified that expression was markedly increased in carcinoma compared with in normal tissue. In conclusion, TBRG4 serves a role in the tumorigenesis of lung cancer via deregulation of DDIT3, CAV1 and RRM2. The results of the present study may be important in contributing to our understanding of TBRG4 as a target for lung cancer treatment.

[Tuberculosis after organ transplantation: clinical analysis and literature review].

OBJECTIVE: To describe the clinical features, diagnosis and treatment of tuberculosis after organ transplantation. METHOD: The clinical data of 25 cases of tuberculosis after organ transplantation were retrospectively analyzed. RESULTS: Sixty-eight percent of the patients presented only mild symptoms. Fever, cough, malaise and chest tightness were among the most common manifestations. Pulmonary tuberculosis, pleurisy, miliary lesions, lymph node disease, and kidney tuberculosis were common. Involvement of two organs were present in 72% (18/25) of the patients. Radiology showed soft lesions, miliary nodules, pleural effusion, hilar and mediastinal lymphadenopathy. Bacteriology and histology were used to confirm the diagnosis in most cases. The average time from presentation to diagnosis was 38 d. The effective rate of therapy was 76% (19/25). CONCLUSION: More attention should be paid to tuberculosis after organ transplantation and immunosuppression. Prompt diagnosis and treatment are important measures to reduce the mortality.

Reduced RKIP expression levels are associated with frequent non-small cell lung cancer metastasis and STAT3 phosphorylation and activation.

The current study examined the role of Raf kinase inhibitor protein (RKIP) in non-small cell lung cancer (NSCLC) metastasis. A total of 100 patients with NSCLC were recruited following pathological diagnosis in the First Affiliated Hospital of Bengbu Medical College. The patients were classified and statistically analyzed according to their clinicopathological characteristics and tumor-node-metastasis stage. Paired tumor tissue and adjacent non-tumor tissue samples were subject to pathological diagnosis and western blot analysis. Transient transfection and lentivirus particle vector-mediated RKIP overexpression, small interfering RNA-mediated silencing, Transwell assays and immunocytochemistry methods were employed to elucidate the role and underlying mechanisms of RKIP and the Janus kinase/signal transducer and activator of transcription (JAK/STAT) signaling pathway in NSCLC metastasis. Furthermore, in order to examine the in vivo effects of RKIP, recombinant lentivirus particles containing the RKIP gene were administrated in a mouse NSCLC tumor model via tail vein injection. The results revealed reduced RKIP expression levels in NSCLC tissue compared with corresponding non-cancer tissue. Additionally, RKIP expression levels were inversely associated with NSCLC intra-lung, lymph node and long-distance metastasis. The results also indicated that RKIP was able to block STAT3 activation via phosphorylation and inhibit NSCLC-cell metastasis in vitro. Furthermore, RKIP knockdown was able to promote STAT3 phosphorylation and cell metastasis in NSCLC cell lines. During in vivo experiments, RKIP overexpression was able to suppress xenograft tumor metastasis in nude mice. Therefore, RKIP may be an important factor in cancer cell metastasis in patients with NSCLC, and RKIP may inhibit NSCLC-cell invasion by blocking the activation of the JAK/STAT3 signaling pathway.

[Clinical application and evaluation of the detection of five drugs resistance genes in Mycobacterium tuberculosis].

OBJECTIVE: To evaluate the clinical application of the detection of the drug resistant genes rPOB, katG, rPSL, PncA and embB in M. tuberculosis by using polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) analysis. METHODS: Gene mutations and anti-tuberculous drug susceptibility test were analyzed in 109 M. tuberculosis isolates by PCR-SSCP and the proportion method on Lowenstein-Jensen medium, respectively. The therapeutic effect was evaluated in patients with tuberculosis. RESULTS: Isolates from more than 50% of the patients with pulmonary tuberculosis were resistant to at least two drugs. The total rates of drug resistance to rifampin (RFP), isoniazid (INH), streptomycin (SM), pyrizinamide (PZA) and ethambutol (EB) were 80.7%, 71.5%, 78.8%, 57.7%, and 48.6%, respectively. The gene mutation rates of rpoB, katG, rpsL, pncA and embB were 76%, 68%, 71%, 51% and 30%, respectively. The gene mutations were correlated with the degree of drug-resistance to M. tuberculosis. Most of the gene mutations were found in drug-resistant isolates in high concentrations. The six month cure rates of MDR-TB, confirmed by drug susceptibility test and by PCR-SSCP, were 54.8% and 62.8%, respectively (P > 0.05). CONCLUSIONS: PCR-SSCP is a sensitive and specific method for rapid detection of rpoB, katG, rpsL, pncA and embB gene mutations in M. tuberculosis. Drug resistant gene detection may be clinically useful in the therapy of tuberculosis.

Hypoxia inducible factor-1α expression is associated with hippocampal apoptosis during epileptogenesis.

Cell apoptosis can cause hippocampal neuronal loss after epileptic seizures. Hypoxia inducible factor (HIF)-1α is an important factor mediating apoptosis after brain injuries, such as cerebral ischemia and traumatic brain injures, but little research has been done on its role in the lithium chloride-pilocarpine induced epileptic model. Here, we used a rat model of pilocarpine-induced status epilepticus (SE) to investigate HIF-1α expression and apoptosis in the hippocampus, and to explore their relationship during epileptogenesis. 120 male Sprague Dawley (SD) rats were treated with lithium chloride-pilocarpine injections and divided into an experimental group (administered by MK-801) and a positive control group (administered by saline). Then the HIF-1α expression and hippocampal apoptosis were investigated by histological confirmation and western blotting at 24h, 3d, 7d and 14d, respectively. The results showed that the administration of MK-801 significantly reduced (P<0.05) HIF-1α expression and hippocampal apoptosis during epileptogenesis in comparison with the positive control. Moreover, the expression of HIF-1α and hippocampal apoptosis presented significant time-dependent changes (P<0.01) within 2 weeks, and their positive correlation (P<0.05) analyzed by Pearson׳s correlation analysis. Meanwhile, the HIF-1α immunostained cells were distributed in accord with TUNEL immunostained cells and Caspase-3 immunopositive cells in the hippocampus. These results indicate that the HIF-1α expression is associated with hippocampal apoptosis, and suggest that HIF-1α is an important factor during epileptogenesis.

Clinical evaluation of a homemade enzyme-linked immunospot assay for the diagnosis of active tuberculosis in China.

The rapid diagnosis of smear-negative pulmonary tuberculosis (TB) and extrapulmonary TB is a significant problem in clinical practice. We evaluated the usefulness of a homemade enzyme-linked immunospot (ELISPOT) assay for the diagnosis of active TB in China. Seventy-eight healthy volunteers, 60 patients with active TB, and 32 patients with non-TB diseases were evaluated by tuberculin skin test (TST), an ELISPOT assay using a recombinant CFP-10/ESAT-6 fusion protein (rCFP-10/ESAT-6) as a stimulant, and T-SPOT-TB assay. The spot-forming cells (SFC) from 78 healthy subjects containing both PPD-positive and -negative persons was 3.7 ± 6.5. Among 31 diagnosed TB patients, the ELISPOT assay had a sensitivity of 67.7%, compared to a sensitivity of 77.4% for the T-SPOT-TB assay. The ELISPOT assay was more sensitive in smear-positive TB cases (76.9%) than in smear-negative TB cases (61.1%), while T-SPOT-TB had roughly similar sensitivities in smear-positive (76.9%) and smear-negative TB cases (77.8%). The specificity was 90.6% for ELISPOT and 78.1% for T-SPOT-TB among 32 subjects with non-TB diseases. The SFC of TB cases was significantly higher than that of non-TB disease cases, and the SFC of smear-positive TB cases was significantly higher than that of smear-negative TB cases (P < 0.01). We confirmed that the homemade ELISPOT assay appears more specific for the diagnosis of active TB than T-SPOT-TB. ELISPOT assay may be a useful method for the rapid diagnosis of active TB, especially for cases of smear-negative TB.