ObjectiveTo study the effect of Tanreqing injection （TRQ） on the pulmonary flora in the rat model of chronic obstructive pulmonary disease （COPD）. MethodWistar rats were randomized into control， model， and TRQ groups. The rats in other groups except the control group were treated by smoking combined with intratracheal instillation of lipopolysaccharide for the modeling of COPD. The TRQ group was intraperitoneally injected with TRQ （2 g·kg^-1）. At the end of the experiment， after blood collection from the abdominal aorta of the rats， the lung tissue was collected for hematoxylin-eosin and picric sirius red staining to reveal the pathological changes. The lung lavage fluid was collected， and the diversity and relative abundance of lung flora in different groups were analyzed by 16S rDNA amplicon sequencing. ResultThe lungs of the control group were normal， and those of the model group showed neutrophil infiltration， telangiectasia， lung hemorrhage and emphysema in individual cases， and thickening of collagen fibers in the trachea. Compared with the model group， the TRQ group showed significantly improved lungs and recovered collagen fibers. The MLI analysis showed that compared with the control group， the model group showcased increased alveolar space （P<0.01）， which was reduced in the TRQ group （P<0.05）. Compared with the control group， the model group showed increased wall thickness （P<0.01）， and the increase was attenuated in the TRQ group （P<0.01）. TRQ increased the Simpson index and altered the α diversity of pulmonary flora. The results of principal co-ordinate analysis showed that TRQ changed the β diversity and reduced the β diversity index of pulmonary flora. At the genus level， the model group showed increased relative abundance of g_Bacillus and g_Brevundimonas and decreased relative abundance of g_Pseudomonas， compared with the control group. After treatment with TRQ， the relative abundance of g_Stenotrophomonas increased， and that of g_Bacillus decreased. The LEfSe of differential taxa between groups showed that the modeling increased the relative abundance of g_Lachnospiraceae_NK4A136_group， and TRQ treatment increased the relative abundance of g_Rhodococcus and g_Stenotrophomonas. ConclusionTRQ can regulate the diversity of pulmonary flora and restore the balance of bacterial genera in the rat model of COPD， which may be one of the mechanisms of the prevention and treatment of COPD with TRQ.

Epidemiological and animal studies indicate that pre-existing diabetes increases the risk of Parkinson's disease(PD).However,the mechanisms underlying this association remain unclear.In the present study,we found that high glucose(HG)levels in the cerebrospinal fluid(CSF)of diabetic rats might enhance the effect of a subthreshold dose of the neurotoxin 6-hydroxydopamine(6-OHDA)on the development of motor disorders,and the damage to the nigrostriatal dopaminergic neuronal pathway.In vitro,HG promoted the 6-OHDA-induced apoptosis in PC12 cells differentiated to neurons with nerve growth factor(NGF)(NGF-PC12).Metabolomics showed that HG promoted hyperglycolysis in neurons and impaired tricarboxylic acid cycle(TCA cycle)activity,which was closely related to abnormal mito-chondrial fusion,thus resulting in mitochondrial loss.Interestingly,HG-induced upregulation of pyruvate kinase M2(PKM2)combined with 6-OHDA exposure not only mediated glycolysis but also promoted abnormal mitochondrial fusion by upregulating the expression of MFN2 in NGF-PC12 cells.In addition,we found that PKM2 knockdown rescued the abnormal mitochondrial fusion and cell apoptosis induced by HG+6-OHDA.Furthermore,we found that shikonin(SK),an inhibitor of PKM2,restored the mito-chondrial number,promoted TCA cycle activity,reversed hyperglycolysis,enhanced the tolerance of cultured neurons to 6-OHDA,and reduced the risk of PD in diabetic rats.Overall,our results indicate that diabetes promotes hyperglycolysis and abnormal mitochondrial fusion in neurons through the upre-gulation of PKM2,leading to an increase in the vulnerability of dopaminergic neurons to 6-OHDA.Thus,the inhibition of PKM2 and restoration of mitochondrial metabolic homeostasis/pathways may prevent the occurrence and development of diabetic PD.

Objective:To investigate the effect of multi line therapy for lung adenocarcinoma transformed into small cell lung cancer (SCLC), and review and discuss the related literature.Methods:Combined with the clinical examples of lung adenocarcinoma transformed SCLC after treatment with anti epidermal growth factor receptor tyrosine kinase inhibitors (EGFR TKIs), the diagnostic process and multi line treatment plan of transformed SCLC were analyzed, and the therapeutic effect was evaluated by imaging. At the same time, it was reviewed and discussed in combination with relevant literature.Results:Serological tumor markers were significant for the diagnosis of transformed SCLC after EGFR-TKI treatment of lung adenocarcinoma, and pathology was still the gold standard for its diagnosis. The multiline therapy of SCLC has certain effect on transformed small cell lung cancer.Conclusion:The overall prognosis of lung adenocarcinoma transformed into SCLC after EGFR TKIs treatment is poor, so it is necessary to diagnose and treat it as early as possible, evaluate the effect of imaging in time, and make treatment adjustment quickly.

Zn²⁺ is required for the activity of many mitochondrial proteins, which regulate mitochondrial dynamics, apoptosis and mitophagy. However, it is not understood how the proper mitochondrial Zn²⁺ level is achieved to maintain mitochondrial homeostasis. Using Caenorhabditis elegans, we reveal here that a pair of mitochondrion-localized transporters controls the mitochondrial level of Zn²⁺. We demonstrate that SLC-30A9/ZnT9 is a mitochondrial Zn²⁺ exporter. Loss of SLC-30A9 leads to mitochondrial Zn²⁺ accumulation, which damages mitochondria, impairs animal development and shortens the life span. We further identify SLC-25A25/SCaMC-2 as an important regulator of mitochondrial Zn²⁺ import. Loss of SLC-25A25 suppresses the abnormal mitochondrial Zn²⁺ accumulation and defective mitochondrial structure and functions caused by loss of SLC-30A9. Moreover, we reveal that the endoplasmic reticulum contains the Zn²⁺ pool from which mitochondrial Zn²⁺ is imported. These findings establish the molecular basis for controlling the correct mitochondrial Zn²⁺ levels for normal mitochondrial structure and functions.
Animals ; Caenorhabditis elegans/metabolism* ; Cation Transport Proteins/genetics* ; Homeostasis ; Mitochondria/metabolism* ; Zinc/metabolism*

Objective    To summarize the clinical experience of surgical resection of synchronous multiple ground-glass nodules (SMGN), and explore the individualized diagnosis and treatment strategy of SMGN. Methods    Clinical data of 84 patients with SMGN who underwent thoracic surgery in Anhui Chest Hospital from July 2016 to August 2018 were analyzed retrospectively, including 18 males (21.4%) and 66 females (78.6%), aged 32-80 (55.6±10.3) years. The results of operation and the information of GGNs were analyzed. Results    Except for 1 patient who was converted to thoracotomy due to extensive dense adhesion of thoracic, other patients underwent video-assisted thoracoscopic surgery successfully. All patients recuperated successfully after operation, without severe perioperative complications or death. Finally, 79 patients were diagnosed as malignant tumors (94.0%), and 5 patients of benign lesions (6.0%). A total of 240 GGNs were removed, among which there were 168 pGGNs, including 68 benign lesions (40.5%) and 100 malignant tumors (59.5%), and 72 mGGNs, including 2 benign nodules (2.8%) and 70 malignant tumors (97.2%). Nodules diameter (P<0.001), consolidation/maximum diameter of nodule ratio (P<0.001), vacuole sign (P<0.001), air bronchograms sign (P=0.001), spine-like process (P=0.001), pleural indentation sign (P<0.001), lobulation sign (P<0.001), and vascular convergence (P=0.002) were correlated with malignant tumor. Conclusion    Analysis of the imaging features of GGNs by thin-section CT scan and three-dimensional reconstruction is of great value in predicting the benign and malignant nodules, which can guide the surgical decision-making and preoperative planning. Through reasonable preoperative planning and following certain principles, simultaneous surgical treatment for SMGN is safe and feasible.

Objective To investigate the effects of leptin on Treg cells and the possible mecha-nism. Methods Leptin-deficient ( ob/ob) mice and homologous wild-type mice were used in this study. The percentages of Treg cells in spleen tissues and peripheral blood samples were measured by flow cytometry ( FCM) . Differences in Treg cell functionality were compared between the two groups. Splenic CD4+T cells, separated from the ob/ob mice and the wild-type mice by magnetic beads, were respectively cultured with leptin and anti-leptin neutralization antibody to evaluate the effects of leptin on Treg cells. Quantitative real-time PCR was performed to analyze the expression of Treg cell-related cytokines at transcriptional level. The levels of IL-10 and TGF-β in the supernatants of CD4+T cell culture were measured with Luminex technolo-gy. Results Compared with the wild-type mice, the ob/ob mice showed higher percentages of Treg cells in both peripheral blood samples and spleen tissues [(11. 56 ± 0. 72)％ vs (5. 47 ± 0. 81)％, (10. 16 ± 0.93)％ vs (6.29±0. 69)％]. Treg cells isolated from the ob/ob mice had stronger immunosuppressive effects on the proliferation of effector T ( Teff) cells and the secretion of TNF-α and IFN-γ than those from the wild-type mice [TNF-α:(1. 6±0. 2)％ vs (2. 4±0. 5)％, IFN-γ:(4. 3±0. 3)％ vs (7. 2±1. 2)％]. The percentages of Treg cells were decreased from (12. 2±1. 8)％ to (7. 6±0. 9)％ upon the in vitro treat-ment of CD4+ T cells from the ob/ob mice with leptin and the immunosuppressive effects of Treg cells were also weakened. However, the percentages of Treg cells were increased from (7. 8±0. 85)％ to (13. 1± 1. 5)％ upon the in vitro treatment of CD4+T cells from the wild-type mice with anti-leptin antibody and the immunosuppressive effects of Treg cells were improved as well. Moreover, the expression of Foxp3, IL-10 and TGF-β at transcriptional level and the levels of IL-10 and TGF-β in the ob/ob group were higher than those in the wild-type group. Conclusions Leptin deficiency significantly promoted the generation of Treg cells in mice and resulted in an increased expression of Foxp3, IL-10 and TGF-βat mRNA level and elevat-ed levels of IL-10 and TGF-β. The treatment of CD4+T cells with leptin might inhibit the generation of Treg cells through down-regulating the transcription of Foxp3, IL-10 and TGF-β.

Objective To investigate the effects of leptin on Th17 cells and the possible mechanism. Methods The leptin-deficient ( ob/ob) mice and their homologous wild-type mice were used in the study.The percentages of Th17 cells in peripheral blood samples, spleen tissues and lymph nodes were measured by flow cytometry ( FCM) analysis.The splenic CD4+T cells, separated from the ob/ob mice and the wild-type mice by using magnetic beads,were respectively cultured with leptin at various concentrations and with anti-leptin neu-tralization antibody to evaluate the effects of leptin on Th17 cells.The quantitative real-time PCR was performed to analyze Th17 cell-related cytokines at transcriptional levels.The levels of IL-6 and IL-17A in the supernatants of CD4+T cell culture were measured with Luminex technology.Results Compared with the wild-type mice, the ob/ob mice showed lower percentages of Th17 cells in both peripheral blood samples and spleen tissues (0.49%±0.03%vs 1.29%±0.1%, 1.56%±0.22%vs 2.47%±0.11%).There was a decrease in the percentages of Th17 cells upon the in vitro treatment of CD4+T cells from wild-type mice with anti-leptin antibody.The per-centages of Th17 cells were increased in a dose-dependent manner upon the in vitro treatment of CD4+T cells from ob/ob mice with leptin.Moreover, the levels of IL-17A and IL-6 and the transcriptional levels of RORγt, IL-17A and IL-6 in leptin deficiency group were lower than those of wild-type group, but were increased upon the treatment with leptin.No significant difference with the transcriptional levels of TGF-βand IL-23 was ob-served between the two groups with and without intervention.Conclusion Leptin deficiency seriously hampered the generation of Th17 cells in mice and resulted in a decreased expression of RORγt, IL-17A and IL-6 at mRNA level.The treatment of CD4 T cells with leptin might promote the generation of Th17 cells through up-regulating the transcription of RORγt and IL-6.

Objective To explore the value of spectral imaging technique in dual-energy CT in differential diagnosis of the metastatic lymph nodes in thyroid carcinoma,squamous cell carcinoma metastatic lymph nodes and lymphoma in the neck.Methods In 30 patients with pathologically confirmed with a total of 79 cervical lymph nodes enlargement which were using dual energy scan .Then observed the change trend of the spectrum curve and comparison the three kinds of lymph node energy spectrum curve’s slope.Results In the 79 lymph nodes,the metastatic lymph nodes in thyroid carcinoma were twenty-three,squamous cell carcinoma metastatic lymph nodes were twenty-four and lymphoma were thirty-two.From 60 to 180 keV,with the increase of keV values,the three kinds of malignant lymph nodes of the corresponding CT value decreasing and the higher the keV value,the CT value decrease magnitude was small,and the spectrum curve was〞drop type〞.The slope spectrum curve of the metastatic lymph nodes of thyroid carcinoma in arterial phase and parenchymal phase were maximum,which were 1.23±0.41 and 0.85±0.33,respectively.The slope spectrum curve of lymphoma in arterial phase and parenchymal phase were least,whcih were 0.40±0.16 and 0.47 ±0.09.The slope spectrum curve of the squamous cell carcinoma metastatic lymph nodes in arterial phase and parenchymal phase were 0.88±0.10 and 0.62±0.28.The energy spectrum curve slope of the three kinds of malignant lymph nodes have statistical significance.Conclusion The energy spectrum curve slope of arterial phase and parenchymal phase has some significance lymph node metastasis of in thyroid carcinoma,the metastatic lymph nodes and lymphoma in the neck.

OBJECTIVETo detect the expression levels of co-inhibitory molecules, including CTLA-4, LAG-3, PD-1 and CD39, on CD4⁺ T cells in peripheral blood or tumor tissues from NSCLC patients and to investigate their potential internal relationships with the progression of NSCLC.

METHODSEighty-eight patients including 53 NSCLC, 17 disease control cases and 18 healthy controls were studied. All the peripheral blood and 13 cases of tumor and tumor-adjacent tissues from surgically treated NSCLC patients were obtained. The expression levels of co-inhibitory molecules CTLA-4, LAG-3, PD-1 and CD39 were assayed by flow cytometry (FCM).

RESULTSThe ratios of CD4⁺ CTLA-4⁺ T cells, CD4⁺ LAG-3⁺ T cells, CD4⁺ PD-1⁺ T cells and CD4⁺ CD39⁺ T cells in the peripheral blood of NSCLC patients were (2.49 ± 2.43)%, (2.47 ± 3.50)%, (12.94 ± 5.96)% and (6.78 ± 5.21)%, respectively, the ratio of CD4⁺ CTLA-4⁺ T cells was significantly higher in the peripheral blood of NSCLC patients than that in the disease controls and healthy controls (P < 0.05) . The ratio of CD4(+)PD-1⁺ T cells was also highly raised in the peripheral blood of NSCLC patients than that in the healthy controls (P < 0.05). Further stratified analysis indicated that the ratio of CD4⁺ PD-1⁺ T cells was (13.21 ± 5.96)% in NSCLC patients entering stages III and IV, also significantly increased as compared with that of (11.06 ± 3.42)% in the patients undergoing stages I and II (P < 0.05). More CD4⁺ CTLA-4⁺ T cells, CD4⁺ LAG-3⁺ T cells and CD4⁺ PD-1⁺ T cells were verified in the cancer tissues (5.07 ± 2.11)%, (7.86 ± 3.24)% and (40.20 ± 18.84)%, respectively, than those in their matched peripheral blood (3.13 ± 1.01)%, (2.65 ± 1.48)% and (15.79 ± 5.69)%, (P < 0.05 for all), and especially, CD4⁺ CTLA-4⁺ T cells and CD4⁺ PD-1⁺ T cells were also highly increased than those in matched cancer-adjacent tissues (P < 0.05 for all).

CONCLUSIONSThe increased expression levels of co-inhibitory molecules CTLA-4, LAG-3 and PD-1 on CD4⁺ T cells in peripheral blood and tumor tissues may be one of the mechanisms related to immune escape of tumor cells, acceleration of disease progression and poor prognosis in NSCLC patients.

Antigens, CD ; metabolism ; Apyrase ; metabolism ; CD4-Positive T-Lymphocytes ; metabolism ; CTLA-4 Antigen ; metabolism ; Carcinoma, Non-Small-Cell Lung ; diagnosis ; metabolism ; Disease Progression ; Flow Cytometry ; Humans ; Lung Neoplasms ; diagnosis ; metabolism ; Programmed Cell Death 1 Receptor ; metabolism

Leptin system plays an important role in lung inlfammation and tumorigenesis, but the precise function in the tumormicrovironment and the prognosis value of the leptin system in lung cancer have not been fully elucidated. hTis re-view will focus on the relationship between leptin system and non-small cell lung cancer (NSCLC), in which special attentions will be paid on the expression of leption and its receptor (LepR) in peripheral blood and tumor tissues, leptin-related signal transduction pathways, the interaction between leption and regulatory T cells (Treg) and the gene polymorphisms of leptin and leptin receptor, and possibly provide new strategies for diagnosis and treatment of NSCLC.