Incidence and risk factors of tuberculosis among the elderly population in China: a prospective cohort study.

BACKGROUND: China is facing challenges of the shifting presentation of tuberculosis (TB) from younger to elderly due to an ageing population, longer life expectancy and reactivation disease. However, the burden of elderly TB and influence factors are not yet clear. To fill the gap, we generated a cohort study to measure the magnitude of TB incidence and associated factors among the elderly population aged 65 years and above in China. METHODS: In this cohort established in 2013 through a prevalence survey conducted in selected sites, a total of 34 076 elderlies without TB were enrolled into two-year follow-up. We used both active and passive case findings to find out all TB patients among them. The person-year (PY) incidence rates for both bacteriologically positive TB and active TB were calculated. Cox proportional regression model was performed to test effect of risk factors, and the population attributable fraction (PAF) of each risk factor contributing to incident TB among elderlies was calculated. RESULTS: Over the two-year follow-up period, a total of 215 incident active TB were identified, 62 of which were bacteriologically positive. The incidence rates for active TB and bacteriologically positive TB were 481.8 per 100 000 PY (95% CI: 417.4-546.2 per 100 000 PY) and 138.9 per 100 000 PY (95% CI: 104.4-173.5 per 100 000 PY), respectively. Incident cases detected by active case finding were significantly higher (P < 0.001). Male, non-Han nationality, previously treated TB, ex/current smoker and body mass index (BMI) < 18.5 presented as independent predictors for developing TB disease. For developing bacteriologically positive TB, the biggest contribution was from self-reported ex or current smoker (18.06%). And, for developing active TB, the biggest contribution was from non-Han nationality (35.40%), followed by male (26.80%) and age at 75 years and above (10.85%). CONCLUSIONS: Ageing population in China had a high TB incidence rate and risk to develop TB disease, implying that National TB Program (NTP) needs to prioritize for elderly. Active case finding should be applied capture more active TB cases among this particular population, especially for male, non-Han nationality, and those with identified risk factors.

CX-3543 Promotes Cell Apoptosis through Downregulation of CCAT1 in Colon Cancer Cells.

AIM: Colon cancer-associated transcript-1 (CCAT1), located in the vicinity of transcription factor c-Myc, was first identified in colon cancer. A small-molecule compound CX3543 (Quarfloxin) selectively targeting Myc G-quadruplexes has entered phase II clinical trials for neuroendocrine carcinomas. The aim of the study was to explore the relationship between CX3543, CCAT1, and cell apoptosis in colon cancer cells. METHODS: Semiquantitative PCR was used to detect the relative expression of CCAT1 in colon cancer (CC) tissues and HT29 cell lines. Real-time PCR (RT-PCR) was also used to investigate the expression of CCAT1 and c-Myc after HT29 cells being treated by CX3543 for 24 h. Cell apoptosis assay and cell proliferation assay were conducted in HT29 cells after being treated by CX3543. RESULTS: The results showed that the expression of CCAT1 was remarkably increased in CC tissues and HT29 cells compared to controls. CX3543 treatment reduced the expression of c-Myc and CCAT1 and promoted cell apoptosis and inhibited cell proliferation. After the expression of CCAT1 was inhibited by sh-CCAT1 transfection, the cell apoptosis rate was higher than that of control group. After the cells were treated by CCAT1 overexpression plasmid transfection and CX3543, the cell apoptosis rate was lower than that of control group. In vivo results showed that CX3543 inhibited the xenograft tumor growth of rats through downregulation of CCAT1. CONCLUSION: Our study demonstrated that CX3543 could inhibit the progression of colon cancer by downregulating CCAT1 expression and might be a potential drug for the treatment of colon cancer.

Risk of Immune-Related Pancreatitis in Patients with Solid Tumors Treated with Immune Checkpoint Inhibitors: Systematic Assessment with Meta-Analysis.

We performed a systematic review and meta-analysis to determine the risk of immune-related pancreatitis associated with the treatment by immune checkpoint inhibitors (ICIs) for solid tumors. Eligible studies were selected from multiple databases including phase II/III randomized controlled trials (RCTs) with ICIs in solid tumor patients. The data were analyzed with Stata version 12.0 software. After excluding ineligible studies, a total of 15 clinical trials were considered eligible for the meta-analysis, which included 9099 patients. Compared with chemotherapy or placebo, the risk ratio (RR) for all-grade lipase elevation after CTLA-4 inhibitor treatment was 1.05 (95% confidence interval (CI): 1.01-2.24, p = 0.047). However, the risk for pancreatitis after ICI treatment in any subgroup was not significantly higher than that after control therapy. In addition, compared with ipilimumab/nivolumab alone, the RR for all-grade and high-grade lipase elevation under combination treatment of nivolumab and ipilimumab was 6.43 (95% CI: 1.43-28.99, p = 0.015) and 6.44 (95% CI: 1.39-29.79, p = 0.017), respectively, and the RR for all-grade amylase elevation under combination treatment was 6.08 (95% CI: 1.51-24.44, p = 0.011). Our meta-analysis has demonstrated that both CTLA-4 inhibitors alone and combination treatment of nivolumab and ipilimumab could increase the risk of amylase or lipase elevation, but not significantly increase the risk of pancreatitis when compared with controls.

[Genetic polymorphism of four X chromosome short temden repeats loci in Hebei Han population].

OBJECTIVE: To investigate the polymorphism of loci DXS6800, DXS6797, GATA172D05, DXS986 four loci in Hebei Han population. METHODS: The genome DNA of unrelated individuals,the families and rotten materials were extracted with phenol-chloroform method and Chelex-100 method,respectively. The PCR products were detected by the polyacrylamide gel electrophoresis and DNA sequencing analysis. RESULTS: Among 150 unrelated males and 150 unrelated females from Hebei Han population, 25 alleles were found in the 4 loci. One hundred and thirty-eight haplotypes of the male were detected. The haplotype diversity reached 0.9986. CONCLUSION: The findings provided the polymorphic data of DXS6800, DXS6797, GATA172D05, and DXS986 loci in Hebei Han population. The four loci are relatively abundant in polymorphic information for identification and the obtained data of Hebei Han population can be applied to the X-STR genetic data bank.

[Methylation status of the IL-10 gene promoter in the peripheral blood mononuclear cells of rheumatoid arthritis patients].

Interleukin 10(IL-10), as an immunoregulatory cytokine, plays an important role in rheumatoid arthritis (RA). IL-10 gene silencing is associated with the chromatin remodeling in differentiated Th1 and Th2 cells. To explore the relationship between IL-10 promoter methylation and gene silencing in the pathogenesis of RA, IL-10 mRNA, protein expression and promoter methylation status were analyzed in the peripheral blood mononuclear cells (PBMC) of 34 RA patients and 30 healthy controls by reverse transcriptase-polymerase chain reaction (RT-PCR), enzyme-linked immunosorbent assay (ELISA) and methylation specific polymerase chain reaction (MSP), respectively. The results showed that IL-10 mRNA and protein expression in RA patients seemed to be lower than that in healthy controls, but there was no statistically significant difference (P>0.05). IL-10 promoter was methylated at a frequency of 85.29% in RA cases, which was significantly higher than the percentage in healthy controls (43.33%) (c 2 =12.439, P=0.000). IL-10 promoter methylation and mRNA expression showed a strong negative correlation (r=-0.579, P=0.001). IL-10 promoter methylation, but not mRNA expression, also correlated statistically with the number of arthritic joints. However, there were no statistical correlations between IL-10 promoter methylation (or mRNA expression) and clinical indices of RA, such as the levels of erythrocyte sedimentation rate (ESR), C reactive protein (CRP) and rheumatic factor (RF) or age (P>0.05). These findings suggest that promoter methylation may be a crucial mechanism of IL-10 gene inactivation in RA and IL-10 promoter CpG island hypermethylation might be involved in the occurrence and development of RA.

[Polymorphism of five short tandem repeat loci on chromosome X in Hebei Han population].

OBJECTIVE: To investigate the polymorphism of DXS6801, DXS6809, DXS7423, DXS7424, DXS9902 five loci in Hebei Han population. METHODS: The PCR products were detected by the polyacrylamide gel electrophresis and DNA sequencing analysis. RESULTS: Among 114 irrelative males and 118 irrelative females from Hebei Han population, 31 alleles were found in the 5 loci. One hundred and one haplotypes of the male were detected and the haplotype diversity reached 0.9975. CONCLUSION: The five loci are relatively abundant in polymorphic information for identification and paternity test. And the obtained data of Hebei Han population can be applied to the X-short tandem repeat genetic data bank.

Effect of cholecystokinin octapeptide on tumor necrosis factor alpha transcription and nuclear factor-kappaB activity induced by lipopolysaccharide in rat pulmonary interstitial macrophages.

AIM: To elucidate the anti-inflammatory mechanism of an intestinal neuropeptide, sulfated cholecystokinin octapeptide (sCCK-8), the effects of sCCK-8 on lipopolysaccharide (LPS)-induced tumor necrosis factor alpha (TNF-alpha) mRNA expression and NF-kappaB activity in pulmonary interstitial macrophages (PIMs) were studied. METHODS: PIMs from rat were stimulated with LPS (1 mg.L(-1)) in the presence or absence of sCCK-8 (10(-8)-10(-6)mol.L(-1)) or/and CCK receptor antagonist proglumide (2 mg.L(-1)). The expression of TNF-alpha mRNA was assayed by reverse transcription polymerase chain reaction (RT-PCR) at 3h of the stimulation, and nuclear factor-kappaB (NF-kappaB) binding activity was analyzed by electrophoretic mobility shift assay (EMSA) at 1 h of stimulation. The IkappaBalpha protein level in the cytoplasma at 30 min of the stimulation was detected by Western blot. RESULTS: sCCK-8, at concentrations from 10(-8) mol.L(-1) to 10(-6) mol.L(-1) obviously inhibited LPS-induced TNF-alpha mRNA expression and NF-kappaB binding activity in a dose-dependent manner, P<0.05, P<0.01. Stimulation PIMs with LPS resulted in a reduction of IkappaBalpha protein level, P<0.01, which was elevated by sCCK-8, P<0.05. The effects of sCCK-8 on NF-kappaB activity and IkappaB protein level were attenuated by CCK receptor antagonist proglumide, P<0.01. CONCLUSION: sCCK-8 inhibits LPS-induced TNF-alpha mRNA expression by regulating NF-kappaB activity in rat PIMs, which is mediated through CCK receptors and inhibiting IkappaB-alpha degradation. This represents one of the anti-inflammatory mechanisms of sCCK-8.

Expression and cell-specific localization of cholecystokinin receptors in rat lung.

AIM: To elucidate whether CCK receptors exist in lung tissues and their precise cellular localization in the lung. METHODS: CCK-AR and CCK-BR mRNA expression and cellular distribution in the rat lung were detected by highly sensitive method of in situ reverse transcription-polymerase chain reaction (RT-PCR) and conventional in situ hybridization. RESULTS: CCK-AR and CCK-BR gene positive signals were observed in bronchial epithelial cells, alveolar epithelial cells, pulmonary macrophages and vascular endothelial cells of the rats' lung by in situ RT-PCR. The hybridization signals of CCK-AR were relatively faint. By in situ hybridization, however, only the signals of CCK-BR but not CCK-AR were detected in the lung, and the positive staining was only found in vascular endothelial cells and macrophages. CONCLUSION: CCK-AR and CCK-BR gene were present in pulmonary vascular endothelial cells, macrophages, bronchial epithelial cells and alveolar epithelial cells, which play an important role in mediating the regulatory actions of CCK-8 on these cells.

[Sequence polymorphism of mtDNA HV1, HV2 overlapping fragments and coding region 8430-8673nt in Han population of Hebei province].

OBJECTIVE: To investigate the sequence polymorphism of mtDNA HV1,HV2 overlapping fragments and coding region encompassing position 8430-8673 in Hebei Han population. METHODS: Polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) combined with sequencing method was used to detect the haplotype distribution of mtDNA in 100 Hebei Han individuals. RESULTS: Ninety-one haplotypes were noted in 100 unrelated individuals. The gene diversity is 0.9985 and the random match probability is 0.0115. Compared with the Anderson sequence, 65 sites of different nucleotide sequences were noted, of which 44 sites were previously registered in MITOMAP, 12 sites were not registered and the gene mutations were different from MITOMAP at 9 positions. CONCLUSION: The obtained data suggest that these loci are valuable genetic markers for personal identification and thus could be used as basic data for the forensic application of mtDNA in Hebei province.

[Polymorphic analysis of four Y short tandem repeat loci in Han nationality of Hebei province in China].

OBJECTIVE: To investigate the distribution of the polymorphism of the Y-chromosomal loci DYS438, DYS439, GATA A7.1 and GATA A7.2 among Han population in Hebei province. METHODS: With the use of PCR followed by polyacrylamide gel electrophoresis and silver staining, the allele frequencies of these loci in 164 unrelated men of Han population were investigated. RESULTS: Four, five, five, four alleles were observed at the loci DYS438, DYS439, GATA A7.1 and GATA A7.2 respectively; the frequencies of these alleles ranged from 0.0359 to 0.6587, from 0.0179 to 0.4107, from 0.0122 to 0.4146 and from 0.0476 to 0.5238 respectively; the probability discrimination of these loci were 0.5121, 0.6811, 0.6679 and 0.6327 respectively. Seventy different haplotypes were found at these loci. Thirty-six different haplotypes appeared only once. The power of discrimination of these four loci was 0.9480. CONCLUSION: The results demonstrate that these loci(DYS438, DYS439, GATA A7.1 and GATA A7.2) are good genetic markers with high determination power and can be applied to individual identification, especially in paternity test and the detection of mixed samples.

[Studies of DNA polymorphism at D7S21 locus in Hebei Han population].

To study the polymorphism at D7S21 locus in Hebei Han population, 124 unrelated individuals were detected rapidly by Minisatellite Variant Repeat-Polymerase Chain Reaction (MVR-PCR) and polyacrylamide gradient gel electrophoresis followed by silver staining,and digital codes were obtained. About 36 digital codes were obtained from each individual. No two unrelated individuals shared the same codes. The probability of identity in 36 digital codes was 3.48 x 10(-18). The percentage of three repeat units, a-type, t-type and 0-type was 48.5%, 49.4% and 2.1% respectively. The heterozygosity (H), excluding probability of paternity (EPP)and polymorphism information content (PIC) were 0.9876, 0.9746 and 0.9872 respectively. The results suggested that D7S21 locus has highly polymorphism in Hebei Han population. The method-polyacrylamide gradient gel electrophoresis followed by silver staining was simple,rapid and practical.

Effect of cholecystokinin octapeptide on diacylglycerol-PKC signaling pathway in rat pulmonary interstitial macrophages stimulated by lipopolysaccharide.

AIM: To investigate the effect of cholecystokinin octapeptide (CCK-8) on the diacylglycerol-protein kinase C (DAG-PKC) signaling pathway in rat pulmonary interstitial macrophages (PIM) stimulated by lipopolysaccaride (LPS). METHODS: The PIM from rat lung tissues were isolated using the collagenase digestion method combined with alveolar lavage and pulmonary vessel perfusion. DAG content and PKC activity were measured by radioenzymatic assay. The translocation of PKCzeta was determined by semi-quantitative immunoblot analysis. RESULTS: CCK-8, at high concentrations (1 x 10(-6) - 1 x 10(-5) mol/L), decreased DAG content and inhibited PKC activity and PKCzeta translocation compared with that in rat resting PIM of a control group (P< 0.01). LPS increased DAG content, and promoted PKC activity and PKCzeta translocation (P< 0.01). CCK-8 decreased LPS-induced DAG content and inhibited LPS-induced PKC activity and PKCzeta translocation significantly at 1 x 10(-8) - 1 x 10(-5) mol/L (P< 0.01). This inhibitory effect of CCK-8 could be abrogated partly by proglumide (non-selective CCK receptor antagonist), CR-1409 (selective CCK-A receptor antagonist), and CR-2945 (selective CCK-B receptor antagonist) in a concentration-dependent manner (P< 0.01). CONCLUSION: CCK-8 was a negative modulator of the DAG-PKC signaling pathway in rat resting PIM, which is very important for maintaining body homeostasis. It significantly inhibited LPS-induced DAG content, PKC activity and PKCzeta translocation in a concentration-dependent manner. The CCK receptor, especially the CCK-A receptor, might play a major role in this process.

Effect of lipopolysaccharide on expression and characterization of cholecystokinin receptors in rat pulmonary interstitial macrophages.

AIM: To investigate the effect of lipopolysaccharide (LPS) on the expression and the binding characteristics of cholecystokinin receptors (CCK-R) in rat pulmonary interstitial macrophages (PIMs). METHODS: The PIMs isolated from rat lung tissues were purified by the collagenase digestion method combined with alveolar lavage and pulmonary vessel perfusion. The expression of CCK-R mRNA was detected by RT-PCR and Southern blot analysis and the binding experiments were performed by radioligand binding assay (RBA). RESULTS: CCK-A receptor (CCK-AR) and CCK-B receptor (CCK-BR) mRNA were detected in rat PIMs and their RT-PCR amplified products had a size of approximately 1.37 kb and 480 bp, respectively. The relative expression of CCK-BR mRNA was higher than that of CCK-AR mRNA after incubation with LPS for 0.5, 2, and 6 h. The expression of CCK-R mRNA could be upregulated obviously by LPS. Southern blot analysis of RT-PCR amplified CCK-AR and CCK-BR mRNA products using [gamma-32P]ATP 5'-end-labelled probe showed specific hybridization bands. The specific binding of [3H]CCK-8S to rat PIM membranes was detected in the rats administered with LPS for 48 h, but not in normal rats. Scatchard analysis of the saturation curves suggested the presence of CCK-R with a high affinity (Kd = 0.68 +/- 0.28 nmol/L) and a low binding capacity (Bmax = 32.5 +/- 2.7 fmol/g protein) in rat PIMs. The specific binding of [3H]CCK-8S to rat PIM membranes was inhibited by unlabelled CCK-8S (IC50 = 2.3 +/- 0.8 nmol/L), CCK-AR specific antagonist CR1409 (IC50 = 0.19 +/- 0.06 micromol/L) and CCK-BR specific antagonist CR2945 (IC50 = 3.2 +/- 0.1 nmol/L). CONCLUSION: Two types of functional CCK-AR and CCK-BR existed in rat PIMs and their expression could be upregulated by LPS.

[Changes of iNOS genes expression in liver following ischemia and reperfusion of limbs and their significance in rats].

AIM: To detect the changes of inducible nitric oxide synthase (iNOS) expression in liver following ischemia/reperfusion (I/R) of hindlimbs and to elucidate their significance. METHODS: I/R was established using the occlusion of the femoral arteries for 4 h and reopening for 2-24 h in rats. The expression of iNOS mRNA, and iNOS protein and the nitrotyrosine (NT), a marker of peroxynitrite (ONOO-), in liver tissue were detected with reverse transcription-polymerase chain reaction (RT-PCR) and immunohistochemical technique, respectively. The liver superoxide dismutase (SOD) activity and malondialdehyde (MDA) contents were spectrophotometrically measured. The observation of pathologic changes of liver was made following the inhibition of iNOS by aminoguanidine (AG). RESULTS: Compared with control groups, the relative expression level of iNOS mRNA significantly increased in I/R group. There were more iNOS positive hepatocytes and more NT positive hepatocytes in I/R group than control groups. The contents of MDA markedly increased, while the activity of SOD significantly decreased in I/R group, compared with those in the control groups. The pathologic changes of rat liver became milder in I/R group following the inhibition of iNOS by AG. CONCLUSION: The expressions of iNOS mRNA and protein in liver are significantly upregulated, excess induction of iNOS-NO is contributed to the liver injury during the I/R of hindlimbs.

[Changes of HO-1 genes expression in liver following ischemia/reperfusion of limbs and their significance in rats].

AIM: To detect the changes of inducible heme oxygenase (HO-1) expression in liver following ischemia/reperfusion (I/R) of hindlimbs and to elucidate their significance. METHODS: I/R was established using the occlusion of the femoral arteries for 4h and reopening for 2-24 h in rats. The expression of HO-1 mRNA and HO-1 protein in liver tissue were detected with reverse transcription-polymerase chain reaction (RT-PCR) and immunohistochemical technique, respectively. The observation of pathologic changes of liver was made following the inhibition of HO-1 by zinc protoporphyrin (ZnPP). RESULTS: Compared with control groups, the relative expression level of HO-1 mRNA significantly increased in I/R group. There were more HO-1 positive hepatocytes in I/R group than control groups. The pathologic changes of liver tissue became more severe in I/R + ZnPP group. CONCLUSION: The expressions of HO-1 mRNA and protein in liver tissue are significantly upregulated, induction of HO-1 is involved in protection for hepatocytes during the I/R of hindlimbs.