Screening and identification of differentially expressed long non-coding RNAs in multidrug-resistant tuberculosis.

BACKGROUND: Efforts to eradicate tuberculosis are largely threatened by drug-resistant tuberculosis, particularly, multidrug-resistant tuberculosis (MDR-TB). Screening and identification potential biomarkers for MDR-TB is crucial to diagnose early and reduce the incidence of MDR-TB. METHODS: To screen the differentially expressed long non-coding RNAs in MDR-TB, the lncRNA and mRNA expression profiles in serum derived from healthy controls (HCs), individuals with MDR-TB and drug-sensitive tuberculosis (DS-TB) were analyzed by microarray assay and 10 lncRNAs were randomly selected for further validation by reverse transcription-quantitative real-time PCR(RT-qPCR). The biological functions of differentially expressed mRNAs as well as relationships between genes and signaling pathways were investigated using Gene Ontology (GO) and the Kyoto Encyclopedia of Genes and Genomes (KEGG), respectively. RESULTS: A total of 353 differentially expressed lncRNAs (312 upregulated) and 202 mRNAs (99 upregulated) were found in the MDR-TB group compared to HCs. And compared with the DS-TB group, 442 differentially expressed lncRNAs (115 upregulated) and 190 mRNAs (87 upregulated) were found in the MDR-TB group. The expression levels of lncRNA n335659 were found to differ significantly between each group by RT-qPCR. Compared with DS-TB group, the GO analysis showed that the differential mRNAs were mainly enriched in the processes associated with the detection of the chemical stimulus, the regulation of mRNA metabolic process and neutrophil activation in the MDR-TB group; the KEGG analysis indicated that the differential mRNAs between DS-TB and MDR-TB were mainly enriched in proteasome and Notch signaling pathway, which might reveal a fraction of the mechanism of MDR-TB. The discovery of the serum lncRNA n335659 might serve as a potential biomarker for MDR-TB and Notch signaling pathway provided a new clue for the investigation of the pathological mechanism of MDR-TB.

Integrating serum microRNAs and electronic health records improved the diagnosis of tuberculosis.

BACKGROUND: To verify the differential expression of miR-30c and miR-142-3p between tuberculosis patients and healthy controls and to investigate the performance of microRNA (miRNA) and subsequently models for the diagnosis of tuberculosis (TB). METHODS: We followed up 460 subjects suspected of TB, and finally enrolled 132 patients, including 60 TB patients, 24 non-TB disease controls (TB-DCs), and 48 healthy controls (HCs). The differential expression of miR-30c and miR-142-3p in serum samples of the TB patients, TB-DCs, and HCs were identified by reverse transcription-quantitative real-time PCR. Diagnostic models were developed by analyzing the characteristics of miRNA and electronic health records (EHRs). These models evaluated by the area under the curves (AUC) and calibration curves were presented as nomograms. RESULTS: There were differential expression of miR-30c and miR-142-3p between TB patients and HCs (p < 0.05). Individual miRNA has a limited diagnostic value for TB. However, diagnostic performance has been both significantly improved when we integrated miR-142-3p and ordinary EHRs to develop two models for the diagnosis of tuberculosis. The AUC of the model for distinguishing tuberculosis patients from healthy controls has increased from 0.75 (95% CI: 0.66-0.84) to 0.96 (95% CI: 0.92-0.99) and the model for distinguishing tuberculosis patients from non-TB disease controls has increased from 0.67 (95% CI: 0.55-0.79) to 0.94 (95% CI: 0.89-0.99). CONCLUSIONS: Integrating serum miR-142-3p and EHRs is a good strategy for improving TB diagnosis.

Nontuberculous mycobacterial pulmonary disease and associated risk factors in China: A prospective surveillance study.

BACKGROUND: We aimed to address the knowledge gap that exists regarding the epidemiological, demographic, and clinical characteristics of nontuberculous mycobacterial pulmonary diseases (NTM-PDs) among smear-positive patients with symptoms suggestive of pulmonary tuberculosis (PTB) in China. METHODS: Prospective and national surveillance of NTM-PD was performed from 17 hospitals within the China Nontuberculous Mycobacteria Surveillance Study (CNTMS). Patients were eligible for inclusion if they had positive smears during hospitalization. Sputum specimens were collected for molecular species identification. RESULTS: 6,766 patients with valid results were included, consisting of 6,236 (92.2%) with PTB, 458 (6.8%) with NTM-PD, and 72 (1.0%) with colonization. The proportion of NTM-PD in PTB patients exhibited significant geographic diversity, ranging from 3.2% in the northwest to 9.2% in the south. The most prevalent species was Mycobacterium intracellulare, followed by Mycobacterium abscessus complex. Females, elderly people, and patients with bronchiectasis or COPD are at high risk for developing NTM-PD, while patients with diabetes have a lower risk of NTM-PD when compared with non-diabetic patients. Regarding clinical symptoms, lower rates of persistent cough and weight loss were noted in NTM-PD patients than in PTB patients. CONCLUSIONS: Approximately one-fifteenth of PTB patients are afflicted with nontuberculous mycobacterial infections in China. The prevalence of NTM shows geographic diversity across the country, and it showed a gradual increase from north to south and from west to east. NTM-PD patients are prone to exhibit less severe clinical symptoms than PTB patients, highlighting the importance of raising awareness of NTM diseases to improve decision making on how to best screen, diagnose, and treat NTM in TB-endemic settings.

Synthesis of antibacterial gold nanoparticles with different particle sizes using chlorogenic acid.

This study proposes a strategy for the rapid and simple synthesis of gold nanoparticles (CGA-AuNPs) with different particle sizes using trisodium citrate (TSC) as the first reducing agent and chlorogenic acid (CGA) as the second reducing agent. And the antibacterial activity of CGA-AuNPs with different particle sizes in vitro was checked by measuring the growth curves of Escherichia coli (ATCC 25922) and Staphylococcus aureus (ATCC 25923). The CGA-AuNPs obtained by the analysis of transmission electron microscope (TEM) images and ultraviolet-visible (UV-Vis) spectra were mainly spherical, and the average diameters were 18.94 ± 1.81, 30.42 ± 6.32, 37.86 ± 3.80 and 48.72 ± 6.47 nm, respectively. High-resolution transmission electron microscopy (HRTEM) and selected area electron diffraction (SAED) showed that these nanoparticles were polycrystalline gold structures. Both CGA-AuNPs and CGA have excellent antibacterial activity, and CGA-AuNPs with small particle size has a stronger antibacterial effect than the larger one. UV-Vis absorption spectrum data revealed that the synthesized CGA-AuNPs without adding other stabilizing agent were well maintained even after 26 days. This work provides a special idea to regulate the size of CGA-AuNPs with CGA by chemical synthesis, and the potent antibacterial activity of these CGA-AuNPs may be applied in the field of antibacterial in the future.

Prolonged parasite clearance in a Chinese splenectomized patient with falciparum malaria imported from Nigeria.

BACKGROUND: The spleen plays a pivotal role in the rapid clearance of parasitized red blood cells in patients with falciparum malaria after artemisinin treatment. Prolonged parasite clearance can be found in patients who have had a splenectomy, or those with hemoglobin abnormalities and/or reduced immunity, which are all distinguishable from artemisinin resistance. This paper reports on a case of prolonged parasite clearance in a Chinese splenectomized patient with falciparum malaria imported from Nigeria. CASE PRESENTATION: A 35-year-old Chinese male suffered 2 days of febrile illness after returning to Zhumadian city of Henan province from Nigeria on October 1, 2014. The main symptoms were febrile, including the highest axillary temperature of 40 °C, headache, and chills. A peripheral blood smear showed parasitemia (53 913 asexual parasites/µl) of Plasmodium falciparum. The patient had not used any chemoprophylaxis against malaria in Nigeria when he worked there as a construction worker between 2009 and 2014. The patient had three episodes of malaria in Nigeria and had a splenectomy due to a traffic accident 8 years ago from the time he was admitted to hospital. The patient was orally administrated a total of 320 mg/2.56 g dihydroartemisinin-piperaquine for 2 days and intravenously administrated a total of 3 000 mg artesunate for 18 days. The axillary temperature of the patient ranged between 37.0 and 37.7 °C from Day 0 to Day 3, and blood microscopy revealed falciparum malaria parasitemia (26 674 asexual parasites/µl) on Day 3. The patient was afebrile on Day 4, falciparum malaria parasitemia was continuously present and then gradually decreased on the next days, and was negative on Day 21. The patient was cured and left hospital on Day 24 after no plasmodium falciparum was found in the blood on Day 21 to Day 23. No mutation was found in the K13 propeller gene when compared with the PF3D7_1343700 K13 propeller gene reference sequence. CONCLUSIONS: This is the first reported case in China of prolonged parasite clearance in a splenectomized patient with imported falciparum malaria. Artemisinin resistance should be distinguished when prolonged parasite clearance is found in a malaria patient who has had splenectomy.

[Protein expressions of HDAC1 and DNMT1 in non-small-cell lung cancer and its clinical significance].

OBJECTIVE: To detect the serum protein expressions of histone deacetylase 1 (HDAC1) and DNA methyltransferase 1 (DNMT1) in patients with non-small-cell lung cancer and explore the correlation between their expressions and clinicopathological features. METHODS: Enzyme-linked immunosorbent assay (ELISA) was used to measure the protein expressions of HDAC1 and DNMT1 in 136 lung cancer patients hospitalized at Department of Respiratory Diseases, First Affiliated Hospital,Zhengzhou University from September 2012 to June 2013 (cancer group). And 147 healthy physical examination subjects from Sixth People's Hospital of Zhengzhou were selected as controls and their clinicopathological features analyzed. RESULTS: The protein expressions of HDAC1 and DNMT1 in patients with lung cancer (145 ± 53, 50 ± 11) were higher than those in control group (78 ± 56, 27 ± 6). And there was statistical significance (t = 596.16, 152.64, both P = 0.000) . The protein expressions of HDAC1 and DNMT1 were correlated with gender, smoking history and age (among P < 0.05). There was a positive association between the protein expressions of HDAC1 and DNMT1 in non-small-cell lung cancer (r = 0.525, P = 0.000). Statistical significance existed between the protein expressions of HDAC1 and DNMT1 and histological types or different stages (all P < 0.01). CONCLUSION: The higher protein expressions of HDAC1 and DNMT1 may play an important role in the early process of lung cancer.

[Expression and significance of DNA methyltransferase in sera of patients with lung cancer].

OBJECTIVE: To explore the protein expressions of DNMT1, DNMT3a and DNMT3b in sera of lung cancer patients. METHODS: Enzyme-linked immunosorbent assay (ELISA) method was used to measure the protein expressions of DNMT1, DNMT3a and DNMT3b in 136 lung cancer patients hospitalized at Department of Respiratory Diseases, First Affiliated Hospital, Zhengzhou University during September 2012 to June 2013. And 147 healthy controls were selected from a population of physical examination at Sixth People's Hospital of Zhengzhou. And the relationship was analyzed between protein expressions of DNMT1, DNMT3a and DNMT3b and clinic characteristics of lung cancer. RESULTS: The protein expressions of DNMT1, DNMT3a and DNMT3b in patients with lung cancer (15 ± 10, 997 ± 76 , 302 ± 25) were higher than those of the controls (13 ± 10, 344 ± 93, 108 ± 22). And there were statistical significance (t = 3.28, 62.51, 37.27; P = 0.021, 0.000, 0.000). The results of Logistic regression show that the protein expressions DNMT1, DNMT3a and DNMT3b increased morbidity for lung cancer (χ(2) = 14.811, 26.768, 12.057; P = 0.000, 0.000 0.001), especially so for DNMT1 (OR = 1.545, 95%CI: 1.238-1.928). No correlation existed between the protein expressions of DNMT1, DNMT3a, DNMT3b and histological types or stages (P > 0.05). CONCLUSIONS: The high protein expressions of serum DNMT1, DNMT3a and DNMT3b increase morbidity for lung cancer. And these markers may predict the early occurrence of lung cancer.

Morphologic studies of high fall injuries.

OBJECTIVE: By studying the mode and morphologic character of high fall injuries on the scene, and exploring the injury situation of different heights, different fall ways and postures, to provide a reference for the forensic identification of high fall injury. METHODS: All the high fall cases were statistically analysed according to their gender, age, ground-touching posture, fall height, site and type of the injury. RESULTS: Among 134 high fall cases, 98 were male and 36 were female with the age ranging from 2-71 years (37.6+/-16.9 on average), in which, 10-60 years old group consisted of 110 cases (82%). Most cases fell from windows or roofs (73%) and the touching objects were cement ground or shaft bottom of elevators. Among these cases, head injury was generally serious, followed by chest and abdominal injuries. The morphologic changes depend upon the height, nature, as well as the posture at the point while the body touches the ground. CONCLUSION: Morphologic study of high fall injury assists medicolegal physicians to make correct identifications of the cause and nature of high fall injuries.

Forensic medical study on morphology and formative mechanism of blunt head injury.

OBJECTIVE: To study the patterns and morphologic characteristics of blunt head injury and analyse its formative mechanism in attempt to provide references for medicolegal expertise. METHODS: The statistical analysis was done in terms of gender, age, as well as the nature, pattern, location, and feature of the injuries. RESULTS: Among the 202 cases of head injury-induced death, 124 were male and 78 female with the age ranging from 1-81 years. Death caused by homicide was dominant (106, 52.5%), followed by suicide (49, 24.3%) and accident (44, 21.8%). The majority of suicide-induced death were by falling from height, and traffic crash was responsible for majority of unexpected death cases. The morphology and pathogenesis of the injuries varied according to differences on the mode, magnitude, and orientation of the outside force giving rise to blunt injury as well as the character of vulnerants. CONCLUSION: Studies on the morphology and its formative rationale of blunt head injury will offer easy access to medicolegal expertise on the mode and character of the injury.

A rabbit model of graded primary mechanical injury to brainstem.

OBJECTIVE: To introduce a new animal model of graded mechanical primary brainstem injury (BSI). METHODS: Altogether 45 rabbits were subjected to BSI by type II biological impact machine designed by the Third Military Medical University. The animals were divided into 4 experimental groups (n equal to 10) and 1 control group (n equal to 5) according to different magnitudes of impact pressure imposed on the occipital nodule: Group 1, 500-520 kPa; Group 2, 520-540 kPa; Group 3, 540-560 kPa; Group 4, 560-580 kPa and Group 5, 0 kPa with 20 kPa increase in each grade. The impact depth was a constant 0.5 cm. After injury, the clinical symptoms and signs as well as pathological changes were observed. RESULTS: Rabbits in Group 1 revealed mild physiological reaction of BSI. They had localized cerebral contusion with punctate hemorrhage and subarachnoid hemorrhage (SAH) was limited to the peripheral tissues at the impact area. In Group 2, obvious physiological reaction was observed. Local pathological lesions reached the superficial layer of brainstem tissues; focal hemorrhage and girdle-shaped SAH in basilar pon were observed under microscope. In Group 3, BSI was more severe with a long respiratory depression. Pathological lesions reached the inner portion of brainstem with massive hemorrhage and the whole brainstem was wrapped by subarachnoid hematoma. In Group 4, most rabbits died due to severe BSI. Pathological lesions deepened to the central brainstem with wide pathological change, rapture of the medulla oblongata central canal. Group 5 was the control group, with normal brainstem structure and no lesion observed. CONCLUSION: This model successfully simulates different levels of brainstem mechanical injury and clearly shows the subsequent pathological changes following injury. It takes two external parameters (impact pressure and depth) and has a similar injury mechanism to clinical accelerating BSI. Moreover it is reproducible and stable, thus being be- neficial for exploring pathophysiological mechanism, diagnosis and forensic identification of various degrees of BSI.

Analysis of 86 fatal motorcycle frontal crashes in Chongqing, China.

OBJECTIVE: To analyze the injuries of motorcyclists involved in fatal motorcycle frontal crashes. METHODS: A survey group involving multi-discipline experts was built to randomly collect data on fatal motorcycle frontal collision accidents that occurred in Chongqing during 2006-2010. The sampled information included medical or autopsy reports, blood alcohol concentration (BAC) level, helmet use, accident witness, field sketch as well as field photos. The motorcyclist injuries were scored according to the Abbreviated Injury Scale (AIS) 2005. The involved riders with a BAC level larger than or equal to 20 mg/ml were attributed to alcohol use. Data were processed statistically with nonparametric test via software SPSS 11.0. RESULTS: A total of 86 fatal motorcycle frontal crashes were sampled and further analyzed. The age of motorcyclists enrolled in this investigation showed nominal distribution and the middle-aged (30-39 years) occupied the highest percentage of fatalities. There were only 14 motorcyclists (16.3%) wearing helmets at the moment of collision. And 12.8% of these motorcyclist crashes were attributable to alcohol use. Impact injury was the main fatal cause, accounting for 72% of motorcyclist deaths, followed by tumbling injury (26%) and run-over (2%). Respectively 84%, 22% and 19% of motorcyclists who sustained head, chest and abdominal trauma died. Extremity injury was the most frequently observed injury type. CONCLUSIONS: This investigation is helpful to build accident prevention programs and develop protection devices which may effectively mitigate injuries and prevent deaths following motorcycle frontal collision accidents. Further investigations on motorcycle collision accidents are still needed.