[Causal relationship between relative abundance of gut microbiota and teratozoospermia: A two-sample Mendelian randomization analysis].

OBJECTIVE: To explore the potential causal relationship between gut microbiota and teratozoospermia. METHODS: We searched the database of Genome-Wide Association Study (GWAS) for gut microbiota- and teratozoospermia-related data. We used gut microbiota as an exposure factor, determined the instrumental variables according to the GWAS data on 18 340 participants released by the MiBioGen Alliance, and derived the outcome variables from the European data on teratozoospermia, with a sample size of 85 716, including 915 cases and 209 006 controls. Using inverse-variance weighting (IVW), MR-Egger regression and the weighted median estimator (WME), we performed two-sample Mendelian randomization (MR) analysis on the retrieved data, and estimated the causal relationship between gut microbiota and teratozoospermia based on the ß value. RESULTS: Two-sample MR analysis indicated that the class Erysipelotrichia, family Erysipelotrichaceae, family Streptococcaceae, genus Coprococcusl, genus Ruminococcaceae UCG009, genus Streptococcus, order Clostridialesm and order Erysipelotrichales were causally related with the increased risk, while the family Porphyromonadaceae with the decreased risk of teratozoospermia. CONCLUSION: The class Erysipelotrichia, family Erysipelotrichaceae, family Streptococcaceae, genus Coprococcusl, genus Ruminococcaceae UCG009, genus Streptococcus, order Clostridialesm and order Erysipelotrichales are one of the causes of teratozoospermia, related to the increased risk of the condition, while the family Porphyromonadaceae has a protective effect on sperm morphology, reducing the risk of teratozoospermia.

[The causal relationship between gut microbiota and male reproductive and sexual health].

OBJECTIVE: The purpose of this study was to use the MR method to explore the causal relationship between 211 gut microbiota and male reproductive and sexual health. METHODS: The MiBioGen alliance published genome-wide association study (GWAS) related genetic variation data was used as instrumental variables (IVs) for gut microbiota, and the Finngen biobank GWAS related genetic variation data was used as IVs for male infertility, abnormal sperm, sexual dysfunction, erectile dysfunction, and testicular dysfunction. The inverse variance-weighted (IVW) method was used as the MR analysis method, the results were evaluated according to the odds ratio and 95% confidence interval of the effect measures, and data sensitivity analysis was performed. RESULTS: The results showed that 6 types of gut microbiota were related to male infertility, 12 types were related to abnormal sperm, 5 types were related to sexual dysfunction, 4 types were related to erectile dysfunction, and 4 types were related to testicular dysfunction. And there was no abnormality in the data sensitivity analysis. CONCLUSION: The intestinal microbiota is closely related to male reproductive and sexual health.

N-acetylcysteine alleviated silica-induced lung fibrosis in rats by down-regulation of ROS and mitochondrial apoptosis signaling.

Reactive oxygen species (ROS) is a normal metabolic product of cellular respiration, but too much ROS can induce cell apoptosis. Here, we used N-acetylcysteine (NAC) to inhibit ROS activity to explore the effects of NAC on silica-induced pulmonary fibrosis in rats and provide evidence for study on the mechanism of silicosis. 24 adult male Sprague-Dawley rats weighing 180-220 g were randomly divided into three groups with eight rats in each group. Silicosis model group and NAC group were adopted non-tracheal exposure method of disposable intrapulmonary injection of 50 g/L, silica suspension 1 mL to establish animal silicosis model, NAC group treated with 600 mg/kg NAC by gavage from the right day of modeling, all animals were sacrificed after 28 days. The level of ROS contents and mitochondrial transmembrane potential changes of AM, the mRNA expression level of type I and type III procollagen, cytochrome C, cysteinyl aspartate specific protease-9 and caspase-3 were detected. The severity of pathological changes and pulmonary fibrosis were observed by pathologic specimens. It was showed that ROS contents and MTP changes were lower in the NAC group compared with the silicosis model group, other indexes were lower in the NAC group than the model group, but higher than those of the control group, the degree of lung fibrotic lesions observed from the pathological slices showed the same trend. These data indicated that NAC can reduce ROS content of AM in silica exposure rats, the mitochondrial apoptosis pathway can also be inhibited, the severity of pulmonary fibrosis alleviated as a result.

Single-cell transcriptomics of blood identified IFIT1+ neutrophil subcluster expansion in NTM-PD patients.

OBJECTIVE: Non-tuberculous mycobacterial pulmonary disease (NTM-PD) is caused by an imbalance between pathogens and impaired host immune responses. Mycobacterium avium complex (MAC) and Mycobacterium abscessus (MAB) are the two major pathogens that cause NTM-PD. In this study, we sought to dissect the transcriptomes of peripheral blood immune cells at the single-cell resolution in NTM-PD patients and explore potential clinical markers for NTM-PD diagnosis and treatment. METHODS: Peripheral blood samples were collected from six NTM-PD patients, including three MAB-PD patients, three MAC-PD patients, and two healthy controls. We employed single-cell RNA sequencing (scRNA-seq) to define the transcriptomic landscape at a single-cell resolution. A comprehensive scRNA-seq analysis was performed, and flow cytometry was conducted to validate the results of scRNA-seq. RESULTS: A total of 27,898 cells were analyzed. Nine T-cells, six mononuclear phagocytes (MPs), and four neutrophil subclusters were defined. During NTM infection, naïve T-cells were reduced, and effector T-cells increased. High cytotoxic activities were shown in T-cells of NTM-PD patients. The proportion of inflammatory and activated MPs subclusters was enriched in NTM-PD patients. Among neutrophil subclusters, an IFIT1+ neutrophil subcluster was expanded in NTM-PD compared to healthy controls. This suggests that IFIT1+ neutrophil subcluster might play an important role in host defense against NTM. Functional enrichment analysis of this subcluster suggested that it is related to interferon response. Cell-cell interaction analysis revealed enhanced CXCL8-CXCR1/2 interactions between the IFIT1+ neutrophil subcluster and NK cells, NKT cells, classical mononuclear phagocytes subcluster 1 (classical Mo1), classical mononuclear phagocytes subcluster 2 (classical Mo2) in NTM-PD patients compared to healthy controls. CONCLUSIONS: Our data revealed disease-specific immune cell subclusters and provided potential new targets of NTM-PD. Specific expansion of IFIT1+ neutrophil subclusters and the CXCL8-CXCR1/2 axis may be involved in the pathogenesis of NTM-PD. These insights may have implications for the diagnosis and treatment of NTM-PD.

[The changes and the significance of cellular immune function in peripheral blood of patients with multidrug-resistant and extensively drug-resistant pulmonary tuberculosis].

OBJECTIVE: To explore the expressions and the significance of CD(3)(+)CD(16)(+)CD(56)(+) NKT cells, CD(3)(-)CD(16)(+)CD(56)(+) NK cells and T lymphocyte subsets in peripheral blood of patients with multi-drug resistant (MDR-TB) and extensively drug-resistant (XDR-TB) pulmonary tuberculosis. METHODS: The data of 316 patients with pulmonary tuberculosis hospitalized in Shanghai Pulmonary Hospital from January 2008 to June 2009 were retrospectively analyzed, of whom 119 were newly diagnosed, and 197 were retreated patients. There were 204 males and 112 females, aged from 17 - 88 years, mean (44 ± 16) years. According to the results of drug-resistance, these patients were divided into a MDR group, an XDR group and a sensitive group. There were 146 patients in the MDR group, with 102 males and 44 females, aged from 19 - 84 years, mean (42 ± 16) years. There were 77 patients in the XDR group, with 42 males and 35 females, aged from 18 - 88 years, mean (50 ± 16) years. There were 93 patients in the susceptible group, with 60 males and 33 females, aged from 17 - 83 years, mean (43 ± 19) years. According to the distribution of cavitation in lung fields, these patients were also divided into 1 - 2 lung field affected group (n = 132), 3 - 4 lung field affected group (n = 49) and 5 - 6 lung field affected group (n = 9). The frequencies of NKT cells, NK cells and T cells from whole blood were tested by flow cytometry. Rank test (SAS software) was used for statistic analyses. RESULTS: The expression rate of NKT cells and NK cells was the highest in the XDR group [11% (6% - 16%) and 7% (4% - 12%)], as compared to the MDR group [8% (5% - 14%) and 6% (4% - 11%)], and the susceptible group [7% (4% - 11%) and 5% (3% - 9%)], the difference being statistically significant (H = 6.478 - 8.369, P < 0.05). The expression rate of the NKT (14 ± 9)% and NK cells (11 ± 7)% in males of the XDR group was significantly higher than that in females [NKT (9 ± 5)% and NK cell (6 ± 4)%], while CD(4) (38 ± 10)% and CD(4)/CD(8) (1.9 ± 1.3) were significantly lower than those of the females [CD(4) (44 ± 10)% and CD(4)/CD(8)(2.2 ± 0.7)], the difference being statistically significant (z = -2.91 - -2.79, P < 0.05, P < 0.01). The expression rate of CD(4) (42 ± 9)% was the highest, but CD(8) (22 ± 8)% was the lowest in the 1 - 2 lung field group. While in the 5 - 6 lung field group, the expression rate of CD(4) (36 ± 11)% was the lowest, CD(8) (28 ± 12)% was the highest, and CD(4)/CD(8) (1.5 ± 0.8) was the lowest, the difference being statistically significant (H = 8.404 - 16.175, P < 0.01). CONCLUSIONS: With the increasing level of drug resistance, the expression rate of NKT cells and NK cells increased, while the expression of T cell subsets did not change. The value of CD(4) and CD(4)/CD(8) in peripheral blood decreased, but CD(8) increased as the extent of cavitation increased in these patients. The impairment of cellular immune function in XDR-TB was more prominent in male patients.

[Effect of SiO2 on expression of MMP-9/TIMP-1 in human alveolar macrophages in vitro].

OBJECTIVE: To investigate the effect of SiO2 on the expression of matrix metalloproteinase-9 (MMP-9) and tissue inhibitor of metalloproteinase-1 (TIMP-1) in human silicotic alveolar macrophages (AM). METHODS: Human alveolar macrophages were collected from a silicotic patient by bronchoalveolar lavage and exposed to silicon dioxide for 3 h, 6 h, 12 h, 18 h, 24 h and 36 h. The expression of the MMP-9 in the AM were detected with zymography and immunological method and the expression of the TIMP-1 in the AM with immunological method. RESULTS: The expressions of MMP-9 in the AM increased clearly along with the time, reached peak at 24 h when detected with zymography (average optical density: 3.061+/-0.153 vs 2.851+/-0.164, P<0.05); and reached peak at 18h when detected with immunological method (average optical density: 0.386+/-0.037 vs 0.322+/-0.034, P<0.05). The expression of the TIMP-1 in the AM did not vary when detected with immunological method (P>0.05). CONCLUSION: SiO2 may affect the expression of MMP-9 and MMP-9 activity in the cultured AM.

[Effect of SiO(2) on expression of platelet derived growth factor in human silicotic alveolar macrophages and human embryonic lung fibroblasts].

OBJECTIVE: To study the effect of SiO(2) on the expression of platelet derived growth factor (PDGF) in human silicotic alveolar macrophages (AM) and human embryonic lung fibroblasts (HELF). METHODS: Human alveolar macrophages were collected from a silicotic patient by bronchoalveolar lavage and exposed to SiO(2) for 3, 6, 12, 18, 24 and 36 h. The cultured supernatant at 24 h was incubated with human embryonic lung fibroblasts for 6, 12, 18, 24, 36 and 48 h. The immunocytochemistry and Western blot were used to detect the level of expression of PDGF in lung fibroblasts and their supernatant respectively. (3)H-proline was used to detect the synthesis and secretion of collagen in HELF. RESULTS: The expression of the PDGF in the supernatant of alveolar macrophages exposed to SiO(2) increased significantly and reached the peak at 24 h (average optical density: 0.282 +/- 0.019 vs 0.214 +/- 0.014, P < 0.01) with ELISA. The expression of PDGF in lung fibroblasts and their supernatant increased at different time (6, 12, 18, 24, 36 and 48 h) with immunocytochemistry and Western blot respectively when incubated with the cultured supernatant of silicotic AM exposed to SiO(2). The expression of PDGF was significantly different from the control group (P < 0.05). The synthesis and secretion of collagen in FB were increased markedly when incubated with the cultured supernatant of AM stimulated by SiO(2) compared with the control group. CONCLUSION: SiO(2) may affect the expression of PDGF and synthesis of collagen through AM mediation and participate in the formation of lung fibrosis.

[Effect of human silicotic alveolar macrophages on the expression of the collagen type I in human embryonic lung fibroblasts].

OBJECTIVE: To study the effect of the cultured supernatant of human silicotic alveolar macrophages (AM) on the expression of the collagen type I in human embryonic lung fibroblasts. METHODS: Human alveolar macrophages were collected from a silicotic patient by bronchoalveolar lavage and exposed to silicon dioxide for 18 h. Then the cultured supernatant were used to culture human embryonic lung fibroblasts for 6 h, 12 h, 18 h, 24 h, 36 h, 48 h, 72 h. Then detected collagen anabolism and secretion with (3)H-proline detected the expression of the procollagen type I in the fibroblast with immunological method detected the quantity of collagen Type I in FB supernatant with Western blot. RESULTS: The anabolism and secretion of collagen were increased in cultured supernatant of silicotic AM exposed to SiO(2), Along with the time, the expression of collagen type I increased. In cultured supernatant of silicotic AM exposed to SiO(2), ((3)H-proline: 1096.500 +/- 76.400, 707.000 +/- 62.160, OD: 0.314 +/- 0.011, OD: 14.218 +/- 0.342. CONCLUSION: SiO(2) may affect the expression of collagen through AM mediation and participate in the formation of lung fibrosis.

[Association of surfactant protein D gene polymorphisms at rs3088308 and rs721917 with susceptibility to silicosis].

OBJECTIVE: To investigate the relationship between polymorphisms of surfactant protein D (rs3088308 and rs721917) and the susceptibility to silicosis. METHODS: This case-control study included 125 silicosis patients and 125 individuals exposed to industrial dust but without silicosis (control group), who were strictly matched with the case group for age, gender, work type and cumulative length of dust exposure. The rs3088308 and rs721917 polymorphisms of surfactant protein-D were detected in all the participants using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). RESULTS: The frequencies of T/T, T/A and A/A genotypes of surfactant protein-D rs3088308 locus were 22.2%, 71.2% and 5.6% in the case group, significantly different from the frequencies of 17.6%, 58.4% and 24.0% in the control group, respectively (P<0.05). The frequencies of C/C, C/T and T/T genotypes of rs721917 locus were 17.6%, 56.8% and 25.6% in the case group, similar to the frequencies of 15.2%, 60.0% and 24.8% in the control group, respectively (P>0.05). CONCLUSION: Surfactant protein-D rs3088308 polymorphism is significantly associated with silicosis, and the T allele may be a risk factor for silicosis in individuals exposed to industrial dust.

Interferon-gamma release assays for the diagnosis of extrapulmonary tuberculosis: a systematic review and meta-analysis.

Interferon -gamma release assays (IGRAs) provide a new diagnostic method for Mycobacterium tuberculosis (TB) infection. However, the diagnostic value of IGRAs for extrapulmonary TB (EPTB) has not been clarified. We searched several databases and selected papers with strict inclusion criteria, evaluated the evidence of commercially available IGRAs (QuantiFERON(®) -TB Gold QFT-G or QFT-GIT and T-SPOT(®) .TB) on blood and the tuberculin skin test (TST) using random effects models. Twenty studies with 1711 patients were included. After excluding indeterminate results, pooled sensitivity for the diagnosis of EPTB was 72% [95% confidence interval (CI) 65-79%] for QFT-G or GIT and 90% (95% CI, 86-93%) for T-SPOT; in high-income countries the sensitivity of QFT-G or GIT (79%, 95% CI 72-86%) was much higher than that (29%, 95% CI 14-48%) in low/middle-income countries. Pooled specificity for EPTB was 82% (95% CI 78-87%) for QFT-G or GIT and 68% (95% CI 64-73%) for T-SPOT. Pooled sensitivity of TST from four studies in high-income countries was lower than that of IGRAs. T-SPOT was more sensitive in detecting EPTB than QFT-G or GIT and TST. However, both IGRAs and TST have similar specificity for EPTB. IGRAs have limited value as diagnostic tools to screen and rule out EPTB, especially in low/middle-income countries. The immune status of patients does not affect the diagnostic accuracy of IGRAs for EPTB.

The elements of human cyclin D1 promoter and regulation involved.

Cyclin D1 is a cell cycle machine, a sensor of extracellular signals and plays an important role in G1-S phase progression. The human cyclin D1 promoter contains multiple transcription factor binding sites such as AP-1, NF-қB, E2F, Oct-1, and so on. The extracellular signals functions through the signal transduction pathways converging at the binding sites to active or inhibit the promoter activity and regulate the cell cycle progression. Different signal transduction pathways regulate the promoter at different time to get the correct cell cycle switch. Disorder regulation or special extracellular stimuli can result in cell cycle out of control through the promoter activity regulation. Epigenetic modifications such as DNA methylation and histone acetylation may involved in cyclin D1 transcriptional regulation.

Efficacy and safety of linezolid in the treatment of extensively drug-resistant tuberculosis.

Linezolid is a new antibiotic with activity against Mycobacterium tuberculosis in vitro and in vivo. This study aims to evaluate the efficacy and safety of linezolid in the treatment of extensively drug-resistant tuberculosis (XDR-TB). We used a linezolid-containing regimen in the treatment of 14 XDR-TB patients. Two years of individualized chemotherapy regimens were adopted on the basis of the patients'medication history and the results of drug susceptibility testing. The patients received 600 mg of linezolid twice a day for the first 1-2 months, followed by once a day thereafter. Eleven patients (78.6%) showed significant improvement in clinical symptoms. Chest computed tomography revealed that 10 patients (71.4%) showed cavity closure. Smear conversion and culture conversion were achieved in all 14 patients (100%) with an average of 64 and 63 days, respectively. The exact proportions of serious and minor adverse events determined by linezolid were 21.4% (3/14) and 64.3% (9/14), respectively. These data show that linezolid-containing chemotherapy for the treatment of XDR-TB may significantly improve clinical symptoms, promote lesion absorption and cavity closure, and accelerate sputum conversion. Further, adverse reactions can be tolerated and resolved with suitable intervention.

[Effects of long-term fertilization on paddy soils organic nitrogen, microbial biomass, and microbial functional diversity].

Soil samples were collected from the plow layers at two long-term experiment sites in Xinhua and Ningxiang counties of Hunan Province, China to study the effects of long-term fertilization on organic nitrogen, microbial biomass, and microbial functional diversity of paddy soils. Long-term fertilization showed great effects on the soil N content. Compared with CK, treatments NPK plus manure or straw increased the contents of soil total acid-hydrolysable N and its fractions amino sugar N, amino acid N, and ammonium N. Treatment NPK had no significant effects on soil microbial biomass C and N, but treatments NPK plus manure increased the contents of soil microbial biomass C and N significantly. BIOLOG test showed that treatments NPK plus manure enhanced the carbon utilization efficiency of soil microbes, and improved the functional diversity of soil microbial communities, compared with CK. Long-term different fertilizer treatments resulted in the differences of carbon substrate utilization patterns of soil microbial communities.

[Effects of long-term fertilization on paddy soil organic carbon mineralization].

An incubation test was conducted with the paddy soil samples collected from three national long-term experiment stations in Hunan Province to study the characteristics of organic carbon mineralization under different fertilization treatments and its relationships with organic carbon fractions, i.e., total organic carbon (TOC), microbial biomass carbon (MBC), and water-soluble organic carbon (WSOC). The results showed that in all fertilization treatments, the cumulative amounts of CO2 and CH4 ranged from 448.64 to 1516.77 microg x g(-1) and from 15.60 to 33.34 microg x g(-1), respectively. In the 58 days of incubation, the mineralized carbon accounted for 3.59%-5.57% of TOC. The CO2 production rate was higher in the early phase of incubation, decreased rapidly then, and tended to stable afterwards; while the CH4 production rate had a slow increase first and declined rapidly then. A combined application of chemical fertilizers and manure or straw increased the cumulative amounts of CO2 and CH4 significantly. In all fertilization treatments, the cumulative mineralized C had significant correlations with TOC, MBC and WSOC, but less correlation with its percentage in TOC.