[Advances in the Study of Tumor-associated Macrophages in Lung Cancer].

Lung cancer is one of the malignant tumors with the highest morbidity and mortality in China. Therefore, the research on the treatment of lung cancer is also deepening. At present, there are mainly systemic chemotherapy, targeted therapy for positive driver genes, the application of immune checkpoint inhibitors, anti-tumor angiogenesis therapy and the combination of the different treatment methods mentioned above. The use of these regimens has significantly improved the prognosis of most lung cancer patients, but the prognosis of patients with advanced lung cancer remains unsatisfactory. Recently, more and more attention has been paid to the study of tumor microenvironment (TME). TME consists of immune cells, fibroblasts, vascular endothelial cells and other cellular components as well as related cytokines, which is the basis for the survival and development of tumor cells. As an important immune cell of TME, tumor-associated macrophages (TAMs) refer to macrophages infiltrating in tumor tissues, which can promote tumor cell proliferation, induce tumor immune tolerance, stimulate tumor angiogenesis, and increase the invasion and metastasis ability of tumor cells. Therefore, targeting TAMs has become a hot topic in lung cancer immunotherapy. In this review, the sources, phenotypes, mechanisms of TAMs in lung cancer, as well as future therapeutic targets of TAMs were reviewed to provide reference for optimal treatment of lung cancer.
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Erratum: Study on clinical characteristics of 173 cases of COVID-19 and effect of glucocorticoid on nucleic acid negative conversion.

[This corrects the article on p. 4251 in vol. 13, PMID: 34150012.].

Study on clinical characteristics of 173 cases of COVID-19 and effect of glucocorticoid on nucleic acid negative conversion.

Objective: To study the clinical characteristics, changes in relevant test parameters, time of nucleic acid negative conversion, and effect of glucocorticoid treatment in Wuhan area patients with the novel coronavirus pneumonia (COVID-19). Methods: Data of 173 inpatients at Huoshenshan Hospital from February 10 to March 17, 2020, were analyzed retrospectively. Clinical characteristics, partial test results, and the influence of glucocorticoid therapy on the clinical outcomes of nucleic acid negative conversion and changes in lung CT images were compared. The patients were divided at admission into 4 groups according to the course of disease and glucocorticoid treatment. Differences among the groups were analyzed statistically. Results: The median age of 173 patients was 62 years, and 91.3% were over 40 years old. Underlying diseases occurred in 50.3% of patients, 32.6% had family gatherings, and 24.3% had exposure while shopping or at a hospital. Median times of nucleic acid negative conversion in group A+B (course of disease < 3 weeks) and group C+D (course of disease ≥ 3 weeks) were 23 days and 37 days, respectively (P < 0.05). Other group comparisons, i.e., of A+C with B+D, A with B, or C with D, were not statistically different. One week after reexamination, chest CT lesion area had changed by 52% in group C and 50% in group D (P > 0.05). In some patients, administration of glucocorticoid for more than 4 weeks significantly promoted the reduction of inflammatory shadow in the lung. Conclusion: Most patients hospitalized with COVID-19 in Wuhan were middle-aged and elderly people with underlying diseases and a history of family gatherings. Glucocorticoid therapy did not affect nor prolong the duration of nucleic acid negative conversion. Glucocorticoid therapy could promote improvement of lung lesions within 3 weeks after disease onset. Beyond 3 weeks, the treatment did not promote reduction in lung shadow area, however the density of shadow did decrease.

Infection Control Precautions and Care Delivery in Hemodialysis Unit during Coronavirus Disease 2019 Outbreak: A Case Series.

BACKGROUND: With an estimated basic reproductive number of 3.77, the Coronavirus Disease 2019 (COVID-19) continues to spread. It is urgent to exert adequate efforts for the management of dialysis patients, caregivers, and healthcare personnel (HCP). This study aimed at reporting practical workflow, identification of high-risk or suspected cases of CO-VID-19, and subsequent response measures. METHODS: At the time of the COVID-19 outbreak, precautions and practice protocols were applied in our dialysis units (DUs). This single-center study retrospectively reviewed all high-risk/suspected cases from January 23, 2020, to February 10, 2020. Epidemiological, clinical feature, and detailed data on all cases were recorded. RESULTS: Practical workflow for the clinical management of dialysis patients, caregivers, and HCP was initiated. A total of 6 high-risk/suspected cases were identified. Female gender, older age, presence of cardiovascular disease, diabetes, anuresis, immunocompromised status, hypoalbuminemia, and underweight were noticeable features in these cases. Direct evidence of infection or epidemiological risk was detected in five cases. Close monitoring for temperature and oxygen saturation during hemodialysis sessions may be reasonable. No confirmed COVID-19 cases were reported in our DU, but certain cases showed rapid deterioration due to other critically severe condition needing hospitalization. Portable dialysis machines are of great need to ensure dialysis care provision. CONCLUSIONS: Our study described a practical workflow for patient-centered management during COVID-19 outbreak. Potential risk factors and underlying clinical patterns were reported. Further studies regarding the efficacy of infection control precautions and practice protocols tailored for dialysis settings are warranted.

Comparison in efficacy and safety of forceps biopsy for peripheral lung lesions guided by endobronchial ultrasound-guided sheath (EBUS-GS) and electromagnetic navigation bronchoscopy combined with EBUS (ENB-EBUS).

Endobronchial ultrasound-guided sheath (EBUS-GS) and electromagnetic navigation bronchoscopy combined with EBUS (ENB-EBUS) are two diagnostic methods used to obtain lung tissue for biopsy of peripheral lung lesions. This study retrospectively summarized the case data of patients who underwent EBUS-GS or ENB-EBUS, both procedures performed at the respiratory endoscopy center of Tangdu Hospital, and the study compared the diagnostic efficacy and complications of the two methods. The study included 93 patients who underwent EBUS-GS and 26 who underwent ENB-EBUS. The diagnostic rates of EBUS-GS and ENB-EBUS were 71.1% and 65.4%, respectively, with no statistical difference (P=0.581). Furthermore, 89.2% of patients in the EBUS-GS group were diagnosed with malignant disease, which was significantly higher than 23.5% diagnosed with malignant disease in the ENB-EBUS group (P=0.00). An analysis of the factors influencing the diagnosis rate showed that the diagnosis rate of EBUS-GS in cases with bronchial signs was 82.5%, which was significantly higher than the 42.9% in the cases in the ENB-EBUS group with bronchial signs (P<0.05). An analysis of the complications showed that the incidence of complications in the EBUS-GS group was 8.4%, and the incidence of complications in the ENB-EBUS group was 3.8%, with no statistical difference (P>0.05). Both EBUS-GS and ENB-EBUS can be used for the diagnosis of peripheral pulmonary disease. However, the diagnostic rate of EBUS-GS is significantly higher than ENB-EBUS in cases with bronchial signs associated with the lesion, and the diagnostic rate of ENB-EBUS in cases with no bronchial signs was higher than that of EBUS-GS with no statistical difference.

Evaluation of bronchoalveolar lavage fluid combined with the loop-mediated isothermal amplification assay in lower respiratory tract infections.

The clinical application of the loop-mediated isothermal amplification (LAMP) assay has been problematic because of conflicting results obtained from the LAMP assay and bacterial culture. In order to eliminate the interference of oral microorganisms and more accurately evaluate the diagnostic performance of the LAMP assay, we utilized bronchoalveolar lavage fluid (BALF) as a sample to test whether the LAMP assay and bacteria culture yielded similar results. A total of 1092 BALF samples from patients with suspected lower respiratory tract infections were collected. For each sample, parallel studies using both bacterial culture and the LAMP assay were carried out. We were the first to utilize BALF as a sample to study the consistency between the LAMP assay and bacterial culture results. The present study demonstrated that the positive rate from the LAMP assay was higher than that from bacterial culture, and the two methods had a better consistency than previously reported.

Automated detection and quantification of COVID-19 pneumonia: CT imaging analysis by a deep learning-based software.

BACKGROUND: The novel coronavirus disease 2019 (COVID-19) is an emerging worldwide threat to public health. While chest computed tomography (CT) plays an indispensable role in its diagnosis, the quantification and localization of lesions cannot be accurately assessed manually. We employed deep learning-based software to aid in detection, localization and quantification of COVID-19 pneumonia. METHODS: A total of 2460 RT-PCR tested SARS-CoV-2-positive patients (1250 men and 1210 women; mean age, 57.7 ± 14.0 years (age range, 11-93 years) were retrospectively identified from Huoshenshan Hospital in Wuhan from February 11 to March 16, 2020. Basic clinical characteristics were reviewed. The uAI Intelligent Assistant Analysis System was used to assess the CT scans. RESULTS: CT scans of 2215 patients (90%) showed multiple lesions of which 36 (1%) and 50 patients (2%) had left and right lung infections, respectively (> 50% of each affected lung's volume), while 27 (1%) had total lung infection (> 50% of the total volume of both lungs). Overall, 298 (12%), 778 (32%) and 1300 (53%) patients exhibited pure ground glass opacities (GGOs), GGOs with sub-solid lesions and GGOs with both sub-solid and solid lesions, respectively. Moreover, 2305 (94%) and 71 (3%) patients presented primarily with GGOs and sub-solid lesions, respectively. Elderly patients (≥ 60 years) were more likely to exhibit sub-solid lesions. The generalized linear mixed model showed that the dorsal segment of the right lower lobe was the favoured site of COVID-19 pneumonia. CONCLUSION: Chest CT combined with analysis by the uAI Intelligent Assistant Analysis System can accurately evaluate pneumonia in COVID-19 patients.

Brequinar inhibits enterovirus replication by targeting biosynthesis pathway of pyrimidines.

Infection of human enteroviruses could cause diverse diseases ranging from mild respiratory symptoms to neurological complications, and even death. Currently, no-FDA approved antiviral drug is available for clinical treatment of human enteroviruses infection. Brequinar is an immunosuppressive drug currently being used for the prevention of organ graft rejection. The drug repurposing studies show that Brequinar exhibits potent antiviral activity against diverse viruses, including flaviviruses, alphavirus, rhabdovirus, and influenza viruses. The antiviral effect of Brequinar on human enterovirus infection has not been investigated yet. Here, the in vitro study shows that Brequinar potently inhibited EV71, EV70, and CVB3 replication at 50% inhibitory concentration (IC50) of 82.40 nM, 29.26 nM, and 35.14 nM, respectively. The antiviral activity of Brequinar was reversed by supplement exogenous pyrimidines, indicating that the antiviral effect of Brequinar against enterovirus relies on the inhibition of dihydroorotate dehydrogenase (DHODH) activity, which is responsible for the de novo biosynthesis of pyrimidines. These data extend the antiviral spectrum of Brequinar and indicate that Brequinar could serve as a promising antiviral drug to treat EV71 and other enterovirus infections.

Use of Interferon-γ release assay for the diagnosis of female genital tuberculosis in Northwest China.

BACKGROUND: Female genital tuberculosis (FGTB) is one of the major causes of infertility. However, nonspecific manifestations and the lack of easy access to gold-standard diagnostic test render a diagnostic difficult for FGTB. The objective of this study was to determine T-SPOT.TB (an interferon-γ release assay, IGRA) performance in patients with FGTB. METHODS: A total of 213 female patients with validated T-SPOT.TB results were recruited in this retrospective study. Among which, 103 were confirmed FGTB, and 110 were excluded from tuberculosis (control). Of the confirmed FGTB patients, 52 were confirmed by microbiologically/histopathologically examination, while the remaining 51 were clinically confirmed (successfully responsive to anti-tuberculosis treatment). T-SPOT.TB test was performed in both FGTB and control group during the diagnostic procedure. RESULTS: The overall sensitivity and specificity of T-SPOT.TB were 86.41% and 75.45% respectively. Sensitivity of T-SPOT.TB was significantly higher when compared with conventional tuberculosis diagnostic tests. Moreover, T-SPOT.TB test using pelvic effusion (PE) showed higher sensitivity than using corresponding peripheral blood (PB) (94.44% vs 72.22%, P < 0.001). Mean value of spot forming cells (SFCs) of T-SPOT.TB using PE was significantly higher than that of PB in FGTB group (193 (IQR 105-280) SFCs/2.5 × 105 PEMCs vs 71 (IQR 36-107) SFCs/2.5 × 105 PBMCs, P = 0.01), while this was not detected in control group (11 (IQR 0-22) SFCs/2.5 × 105 PEMCs vs 9 (IQR 0-18) SFCs/2.5 × 105 PBMCs, P = 0.77). CONCLUSION: These results demonstrated that T-SPOT.TB, especially PE T-SPOT.TB, is an useful adjunct in FGTB diagnosis.

Heat-Labile Enterotoxin-Induced PERK-CHOP Pathway Activation Causes Intestinal Epithelial Cell Apoptosis.

Enterotoxigenic Escherichia coli (ETEC) is a leading cause of diarrhea among children and travelers in developing countries, and heat-labile enterotoxin (LT) is one of the most important virulence factors. The pathogenesis of and virulence factors associated with ETEC have been well-characterized; however, the extent to which ETEC damages host cells remains unclear. In this study, we found that LT could induce decreases in intestinal epithelial cell viability and induce apoptosis in a dose- and time- dependent manner in both HCT-8 and Caco-2 cells. We analyzed the expression profiles of apoptosis-related proteins via protein array technology and found that Bax, p-p53(S46), cleaved caspase-3, and TNFRI/TNFRSF1A expression levels were significantly up-regulated in wild-type ETEC- but not in ΔLT ETEC-infected HCT-8 cells. Bax is essential for endoplasmic reticulum (ER) stress-triggered apoptosis, and our RNAi experiments showed that the PERK-eIF2-CHOP pathway and reactive oxygen species (ROS) are also main participants in this process. LT-induced ROS generation was decreased in CHOP-knockdown HCT-8 cells compared to that in control cells. Moreover, pretreatment with the ROS inhibitor NAC down-regulated GRP78, CHOP, Bim, and cleaved caspase-3 expression, resulting in a reduction in the apoptosis rate from 36.2 to 20.3% in LT-treated HCT-8 cells. Furthermore, ROS inhibition also attenuated LT-induced apoptosis in the small intestinal mucosa in the ETEC-inoculation mouse model.

MiR-23a-5p modulates mycobacterial survival and autophagy during mycobacterium tuberculosis infection through TLR2/MyD88/NF-κB pathway by targeting TLR2.

Autophagy plays a pivotal role in activating the antimicrobial host defense against Mycobacterium tuberculosis (M.tb.). The emerging roles of microRNAs (miRNAs) in regulating immune responses have attracted increasing attention in recent years. Appreciating the potential of host-directed therapies designed to control autophagy during mycobacterial infection, we focused on the influence of miR-23a-5p on the activation of macrophage autophagy during M.tb. infection in bone marrow-derived macrophages (BMDMs) and murine RAW264.7 cells. Here, we demonstrated that M.tb.-infection of macrophages lead to markedly enhanced expression of miR-23a-5p in a time- and dose-dependent manner. Furthermore, forced expression of miR-23a-5p accelerated the survival rate of intracellular mycobacteria, while transfection with miR-23a-5p inhibitors attenuated mycobacterial survival. More importantly, overexpression of miR-23a-5p dramatically prevented M.tb.-induced activation of autophagy in macrophages, whereas inhibitors of miR-23a-5p remarkably accelerated M.tb.-induced autophagy. Mechanistically, miR-23a-5p is able to modulate TLR2/MyD88/NF-κB signaling activity by targeting TLR2 in RAW264.7 cells in response to M.tb.-infection. Collectively, these findings demonstrated that miR-23a-5p modulated the innate host defense by promoting mycobacteria survival and inhibiting the activation of autophagy against M.tb. through TLR2/MyD88/NF-κB pathway by targeting TLR2, which may provide a promising therapeutic target for tuberculosis.

Electron Acceptors Induce Secretion of Enterotoxigenic Escherichia coli Heat-Labile Enterotoxin under Anaerobic Conditions through Promotion of GspD Assembly.

Heat-labile enterotoxin (LT), the major virulence factor of enterotoxigenic Escherichia coli (ETEC), can lead to severe diarrhea and promotes ETEC adherence to intestinal epithelial cells. Most previous in vitro studies focused on ETEC pathogenesis were conducted under aerobic conditions, which do not reflect the real situation of ETEC infection because the intestine is anoxic. In this study, the expression and secretion of LT under anaerobic or microaerobic conditions were determined; LT was not efficiently secreted into the supernatant under anaerobic or microaerobic conditions unless terminal electron acceptors (trimethylamine N-oxide dihydrate [TMAO] or nitrate) were available. Furthermore, we found that the restoration effects of TMAO and nitrate on LT secretion could be inhibited by amytal or ΔtorCAD and ΔnarG E. coli strains, indicating that LT secretion under anaerobic conditions was dependent on the integrity of the respiratory chain. At the same time, electron acceptors increase the ATP level of ETEC, but this increase was not the main reason for LT secretion. Subsequently, the relationship between the integrity of the respiratory chain and the function of the type II secretion system was determined. The GspD protein, the secretin of ETEC, was assembled under anaerobic conditions and was accompanied by LT secretion when TMAO or nitrate was added. Our data also demonstrated that TMAO and nitrate could not induce the GspD assembly and LT secretion in ΔtorCAD and ΔnarG strains, respectively. Moreover, GspD assembly under anaerobic conditions was assisted by the pilot protein YghG.

Lewis Lung Cancer Cells Promote SIGNR1(CD209b)-Mediated Macrophages Polarization Induced by IL-4 to Facilitate Immune Evasion.

Tumor-associated macrophages are a prominent component of lung cancer and contribute to tumor progression by facilitating the immune evasion of cancer cells. DC-SIGN (CD209) assists in the immune evasion of a broad spectrum of pathogens and neoplasms by inhibiting the maturation of DCs and subsequent cytokines production. However, the expression of DC-SIGN in macrophages and its role in mediating immune evasion in lung cancer and the underlying mechanism remain unclear. Our study aimed to identify the immunosuppressive role of SIGNR1 in murine macrophage differentiation and lung cancer progression. We found that SIGNR1-positive RAW264.7 macrophages were enriched in mixed cultures with Lewis lung cancer cells (LLC) (ratio of RAW 264.7 to LLC being 1:1) after stimulation with IL-4. Moreover, LLC-educated macrophages exhibited significantly higher levels of IL-10 but lower IL-12 in response to IL-4 treatment as determined by RT-PCR and ELISA. However, inhibition of SIGNR1 markedly hampered the production of IL-10, indicating that SIGNR1 was indispensable for IL-4+LLC induced macrophage polarization towards the M2 subtype. Furthermore, polarized M2 cells immersed in a tumor microenvironment promoted the migration of LLCs, as measured by transwell assays, but migration was suppressed after blockade of SIGNR1 using CD209b antibody. In addition, IL-4+LLC-educated macrophages reduced the proliferation of the activated T cells and reduced IFN-γ-mediated Th1 response in T cells, while SIGNR1 inhibition rescued Th1 cell functions. In conclusion, murine SIGNR1 expressed in LLC-educated macrophages appears to mediate IL-4-induced RAW264.7 macrophage polarization and thus facilitate lung cancer evasion.

SIRT1 protects against myocardial ischemia-reperfusion injury via activating eNOS in diabetic rats.

BACKGROUND: Diabetic patients are more sensitive to myocardial ischemic injury than non-diabetic patients. Silent information regulator 1 (SIRT1) is a nicotinamide adenine dinucleotide-dependent histone deacetylase making the heart more resistant to ischemic injury. As SIRT1 expression is considered to be reduced in diabetic heart, we therefore hypothesized that up-regulation of SIRT1 in the diabetic heart may overcome its increased susceptibility to ischemic injury. METHODS: Male Sprague-Dawley rats were fed with high-fat diet and injected with streptozotocin once to induce diabetes. Diabetic rats received injections of adenoviral vectors encoding SIRT1 (Ad-SIRT1) at five myocardial sites. Four days after adenoviral injection, the rats were subjected to myocardial ischemia and reperfusion (MI/R). Outcome measures included left ventricular function, infarct size, cellular death and oxidative stress. RESULTS: Delivery of Ad-SIRT1 into the hearts of diabetic rats markedly increased SIRT1 expression. Up-regulation of SIRT1 in diabetic hearts improved cardiac function and reduced infarct size to the extent as in non-diabetic animals following MI/R, which was associated with reduced serum creatine kinase-MB, lactate dehydrogenase activities and cardiomyocyte apoptosis. Moreover, Ad-SIRT1 reduced the increase in the superoxide generation and malonaldialdehyde content and simultaneously increased the antioxidant capability. Furthermore, Ad-SIRT1 increased eNOS phosphorylation and reduced eNOS acetylation in diabetic hearts. NOS inhibitor L-NAME inhibited SIRT1-enhanced eNOS phosphorylation, and blunted SIRT1-mediated anti-apoptotic and anti-oxidative effects and cardioprotection. CONCLUSIONS: Overexpression of SIRT1 reduces diabetes-exacerbated MI/R injury and oxidative stress via activating eNOS in diabetic rats. The findings suggest SIRT1 may be a promising novel therapeutic target for diabetic cardiac complications.

DDX5 promotes proliferation and tumorigenesis of non-small-cell lung cancer cells by activating ß-catenin signaling pathway.

The DEAD-box-protein DDX5 is an ATP-dependent RNA helicase that is frequently overexpressed in various cancers and acts as a transcriptional co-activator of several transcription factors, including ß-catenin. DDX5 is reported to be involved in cancer progression by promoting cell proliferation and epithelial-mesenchymal transition. However, the clinical significance and biological role of DDX5 in non-small-cell lung cancer (NSCLC) remain largely unknown. In this study, we examined the expression of DDX5 in clinical NSCLC samples, investigated its role in regulating NSCLC cell proliferation and tumorigenesis, and explored the possible molecular mechanism. We found that DDX5 was significantly overexpressed in NSCLC tissues as compared with the matched normal adjacent tissues. In addition, overexpression of DDX5 was associated with advanced clinical stage, higher Ki67 index, and shorter overall survival in NSCLC patients. Upregulation of DDX5 promoted proliferation of NSCLC cells in vitro and growth of NSCLC xenografts in vivo, whereas downregulation of DDX5 showed the opposite effects. Furthermore, DDX5 directly interacted with ß-catenin, promoted its nuclear translocation, and co-activated the expression of cyclin D1 and c-Myc. ß-catenin silencing significantly abrogated DDX5-induced cyclin D1 and c-Myc expression and proliferation in NSCLC cells. Interestingly, DDX5 and cyclin D1 expression followed positive correlation in the same set of NSCLC samples. These findings indicated that DDX5 played an important role in the proliferation and tumorigenesis of NSCLC cells by activating the ß-catenin signaling pathway. Therefore, DDX5 may serve as a novel prognostic marker and potential therapeutic target in the treatment of NSCLC.

ZC3H12D attenuated inflammation responses by reducing mRNA stability of proinflammatory genes.

Infection in airspaces and lung parenchyma may cause acute lung injury and multiple organ dysfunction syndrome due to acute inflammatory response, leading to organ failure and high mortality. ZC3H12D has been shown to modulate Toll-like receptor signaling. This study aimed to investigate the change of ZC3H12D during acute lung injury and its role in inflammation processes. Mice were challenged with lipopolysaccharides (LPS) intratracheally. The expression levels of Zc3h12d, NF-κB, and cytokines were analyzed by quantitative real-time PCR (qPCR), ELISA, and Western blot. The mRNA stability was assessed by qPCR after cells were treated with actinomycin D for specified times. The 3' untranslated region (3'-UTR) of c-fos was cloned immediately downstream of the luciferase coding sequence driven by CMV promoter and luciferase activity was measured with a Luciferase Assay kit. Upon LPS treatment, ZC3H12D levels were reduced in mouse immune cells, whereas levels of NF-κB, IL-6, and TNF-α were significantly increased. Knockdown Zc3h12d in THP1 cells resulted in the upregulation of NF-κB while overexpression of Zc3h12d inhibited NF-κB expression. Ectopic Zc3h12d significantly reduced the mRNA stability of c-fos, NF-κB, TNF-α, IL-1ß, and IL-6. Attachment of the c-fos 3'-UTR made luciferase expression levels sensitive to levels of ZC3H12D. The data indicated that ZC3H12D could suppress both the initial inflammation storm and chronic inflammation by targeting the mRNA of cytokines as well as NF-κB and c-fos.

Tetraspanin CD63 is a regulator of HIV-1 replication.

Macrophages and CD4(+) T-cells are the major reservoirs for HIV-1 infection. CD63 is a tetraspanin transmembrane protein, which has been shown to play an essential role during HIV-1 replication in macrophages. In this study, we further confirm the requirement of CD63 in HIV-1 replication events in primary human CD4(+) T-cells, dendritic cells, and a CD4(+) cell line. Most interestingly, we also show the evidences for the co-localization and internalization of CD63 and HIV-1 major receptor CD4 in primary human macrophages and CD4(+) cell line by confocal microscopy and Co-Immunoprecipitation assay. Analysis revealed that CD63-depleted CD4(+) T-cells, dendritic cells, and a cell line showed significant decrease in HIV-1 production. Further analysis showed that CD63 down regulation reduced production of the early HIV protein Tat, and affected HIV protein Gag by CD63-Gag interaction. In agreement, CD63 silencing also inhibited production of the late protein p24. Furthermore, we revealed that CD63 silencing has no effect on HIV-1 replication with extensive viral challenge (MOI > 0.2). These findings suggest that CD63 plays a dual-role both in early and late HIV-1 life cycle with a range of HIV-1 infection (MOI < 0.2).

Calorie Restriction Attenuates Monocrotaline-induced Pulmonary Arterial Hypertension in Rats.

Calorie restriction (CR) is one of the most effective nonpharmacological interventions protecting against cardiovascular disease, such as hypertension in the systemic circulation. However, whether CR could attenuate pulmonary arterial hypertension (PAH) is largely unknown. The PAH model was developed by subjecting the rats to a single subcutaneous injection of monocrotaline. CR lowered mean pulmonary arterial pressure (mPAP) and reduced vascular remodeling and right ventricular hypertrophy in PAH rats. Meanwhile, CR attenuated endothelial dysfunction as evidenced by increased relaxation in response to acetylcholine. The beneficial effects of CR were associated with restored sirtuin-1 (SIRT1) expression and endothelial nitric oxide synthase (eNOS) phosphorylation and reduced eNOS acetylation in pulmonary arteries of PAH rats. To further clarify the role of SIRT1 in the protective effects of CR, adenoviral vectors for overexpression of SIRT1 were administered intratracheally at 1 day before monocrotaline injection. Overexpression of SIRT1 exhibited similar beneficial effects on mPAP and endothelial function, and increased eNOS phosphorylation and reduced eNOS acetylation in the absence of CR. Moreover, SIRT1 overexpression attenuated the increase in mPAP in hypoxia-induced PAH animals. Overall, the present data demonstrate that CR may serve as an effective treatment of PAH, and targeting the SIRT1/eNOS pathway may improve treatment of PAH.

Novel bronchoscopic balloon dilation for patients with bronchostenosis caused by bronchial tuberculosis: a case report.

INTRODUCTION: Bronchoscopic balloon dilation is a common method in the treatment of bronchostenosis but it is not an effective treatment due to its short dilating time (3 minutes) and low pressure (<3atm). Until recently, the reported highest dilating pressure was ≤6atm; however, this is not enough pressure to dilate a bronchostenosis because of the resistance of the bronchus. We hypothesized that higher dilating pressure (up to 14atm) with longer dilating time (40 minutes) may make bronchoscopic balloon dilation treatment more effective according to the blood vessel dilating method. Therefore, we designed this new bronchoscopic balloon dilation method for treating bronchostenosis, particularly in cases caused by bronchial tuberculosis. CASE PRESENTATION: A 23-year-old Chinese woman presented with right middle segmental bronchostenosis caused by bronchial tuberculosis. She was informed of the surgical procedure and she provided informed consent. After taking anti-bronchial tuberculosis drugs for 2 months, she underwent our new bronchoscopic balloon dilation treatment (dilating time, 40 minutes; pressure, 14atm). After anti-bronchial tuberculosis treatment for 13 months, her intermediate bronchus was observed with videobronchoscopy again and no re-stenosis was seen. Furthermore, a computed tomography scan revealed that her right lower lobe and right middle lobe had reopened. No complications occurred in the patient. CONCLUSION: The novel high-handed videobronchoscopic balloon dilation method was safe and effective for treating this patient with bronchostenosis caused by bronchial tuberculosis.

Epigallocatechin-3-gallate ameliorates seawater aspiration-induced acute lung injury via regulating inflammatory cytokines and inhibiting JAK/STAT1 pathway in rats.

Signal transducers and activators of transcriptions 1 (STAT1) play an important role in the inflammation process of acute lung injury (ALI). Epigallocatechin-3-gallate (EGCG) exhibits a specific and strong anti-STAT1 activity. Therefore, our study is to explore whether EGCG pretreatment can ameliorate seawater aspiration-induced ALI and its possible mechanisms. We detected the arterial partial pressure of oxygen, lung wet/dry weight ratios, protein content in bronchoalveolar lavage fluid, and the histopathologic and ultrastructure staining of the lung. The levels of IL-1, TNF-α, and IL-10 and the total and the phosphorylated protein level of STAT1, JAK1, and JAK2 were assessed in vitro and in vivo. The results showed that EGCG pretreatment significantly improved hypoxemia and histopathologic changes, alleviated pulmonary edema and lung vascular leak, reduced the production of TNF-α and IL-1, and increased the production of IL-10 in seawater aspiration-induced ALI rats. EGCG also prevented the seawater aspiration-induced increase of TNF-α and IL-1 and decrease of IL-10 in NR8383 cell line. Moreover, EGCG pretreatment reduced the total and the phosphorylated protein level of STAT1 in vivo and in vitro and reduced the phosphorylated protein level of JAK1 and JAK2. The present study demonstrates that EGCG ameliorates seawater aspiration-induced ALI via regulating inflammatory cytokines and inhibiting JAK/STAT1 pathway in rats.