[Microglia differential genes and their functions in paraquat-induced Parkinson's disease-like in mice's brains based on single-cell RNA sequencing].

Objective: To analyze the differential genes and related signaling pathways of microglia subpopulations in Parkinson's disease (PD) -like mouse brains induced by paraquat (PQ) based on single-cell RNA sequencing, and provide clues to elucidate the mechanism of PQ-induced PD-like changes in the brain of animals. Methods: In September 2021, six male 6-week-old C57BL/6 mice were randomly divided into control group and experimental group (three mice in each group) . The mice were injected with saline, 10.0 mg/kg PQ intraperitoneally, once every three days, and 10 consecutive injections were used for modeling. After infection, the brains of mice were taken and 10×Genomics single-cell RNA sequencing was performed. Microglia subpopulations were screened based on gene expression characteristics, and Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses were performed. The differential genes of microglia subpopulations between the experimental group and control group were further screened, and functional enrichment analysis was performed using bioinformatics tools. Mouse microglia (BV2 cells) were treated with 0, 60, 90 µmol/L PQ solution, respectively. And real-time fluorescence quantitative PCR experiments were conducted to validate the expressions of differential genes hexokinase 2 (Hk2) , ATPase H+ Transporting V0 Subunit B (Atp6v0b) and Neuregulin 1 (Nrg1) . Results: Cluster 7 and Cluster 20 were identified as microglia subpopulations based on the signature genes inositol polyphosphate-5-phosphatase d, Inpp5d (Inpp5d) and transforming growth factor beta receptor 1 (Tgfbr1) , and they reflected the microglia-activated M2 phenotype. The bioinformatics analysis showed that the characteristic genes of identified microglia subpopulations were enriched in endocytosis. In terms of molecular function, it mainly enriched in transmembrane receptor protein kinase activity and cytokine binding. The up-regulated genes of Cluster 7 were mainly enriched in lysosomal pathway, endocytosis pathway, and down-regulated genes were mainly enriched in neurodegenerative disease and other signaling pathways. The up-regulated genes of Cluster 20 were mainly enriched in signaling pathways related to PD, and down-regulated genes were mainly enriched in cyclic adenosine 3', 5'-monophosphate (cAMP) signaling pathways, neurological development, synaptic function and other signaling pathways. The results of real-time fluorescence quantitative PCR showed that the expressions of Hk2 mRNA and Atp6v0b mRNA increased and the expression of Nrg1 mRNA decreased in the 90 µmol/L PQ-treated BV2 cells compared with the 0 µmol/L, and the differences were statistically significant (P<0.05) . Conclusion: Microglia are activated in the PQ-induced PD-like mouse model and polarized toward the M2 phenotype. And their functions are associated with lysosomal (endocytosis) , synaptic functions and the regulation of PD-related pathways.

[Expression level of Wilms' tumor 1 gene and its correlation with clinical features in patients with myeloproliferative neoplasms].

Objective: To investigate the expression level of WT1 gene in patients with classical Philadelphia chromosome (Ph)-negative myeloproliferative neoplasms (MPN) and its correlation with clinical features. Methods: A retrospective study included 252 patients with newly diagnosed MPN in Zhongnan Hospital of Wuhan University from January 2015 to March 2023, including 128 males and 124 females, aged［M（Q1，Q3）］62 (53, 69) years. The WT1-positive group (n=93) and the WT1-negative group (n=159) were split based on the level of WT1 gene expression, and the variations in clinical indicators between the two groups were compared. Its levels of expression in each subtype and its relationships to thrombotic events and clinically significant variables were analyzed. As of March 31, 2023, the follow-up period [M (Q1, Q3)] was 12.0(6.5,21.0)months. The risk factors of thrombosis in MPN patients were analyzed by using the logistic regression analysis. Results: The WT1 gene expression level in the overall bone marrow samples of 252 patients with newly diagnosed MPN was 0.30% (0.10%, 1.10%). The expression level in primary myelofibrosis (PMF) patients was 1.45% (0.41%, 3.24%), which was higher than 0.15% (0.02%, 0.32%), 0.37% (0.16%, 1.09%) in essential thrombocythemia (ET) and polycythemia vera (PV) patients (both P<0.05). Positive correlations were found between WT1 gene expression levels and JAK2V617F gene mutation load, RDW, MPV (r=0.478, 0.346, 0.236, all P<0.01). While negative correlations between WT1 gene expression levels and PLT, LYM, PTTA, LDH were found (r=-0.339, -0.170, -0.206, -0.388, all P<0.01). Patients in the WT1-positive group exhibited a higher percentage of somatic symptoms, splenomegaly, positive JAK2V617F gene mutation, and higher levels of RDW, LDH, NEUT, and MPV compared to the WT1-negative group. In contrast, the proportion of triple-negative (negative for all three hot mutations of JAK2V617F, CALR and MPL) was lower, and the levels of PLT, LYM and PTTA were lower (all P<0.05). The thrombotic event rates of WT1-positive group and WT1-negative group were 32.3% (30/93) and 32.1% (51/159), respectively, and the difference was not statistically significant (P=0.883). Logistic regression analysis showed that male (OR=2.41,95%CI:1.02-5.71,P=0.046) and positive JAK2V617F gene mutation (OR=3.96,95%CI:1.50-10.42,P=0.005) were risk factors for thrombotic events in ET patients. Conclusions: WT1 gene expression is elevated in PMF patients and correlated with indicators of disease progression and transformation in MPN patients. It can be utilized as an auxiliary diagnostic indicator for classical MPN staging but is not correlated with the incidence of thrombotic events. Male and positive JAK2V617F gene mutation are risk factors for thrombotic events in ET patients.

[Definition of severe pulmonary tuberculosis: a scoping review].

Objective: To clarify the definition of severe pulmonary tuberculosis and its inclusion criteria by summarizing and analyzing the studies of severe pulmonary tuberculosis (TB). Methods: A systematic search of Medline (via PubMed), Cochrane Library, Web of Science, Web of Science, Epistemonikos, Embase, CNKI, WanFang database, and CBM database was conducted to collect studies published between 2017 and 2022 on patients with severe pulmonary TB. Searches were performed using a combination of subject terms and free words. The search terms included: tuberculosis, severe, serious, intensive care, critical care, respiratory failure, mechanical ventilation, hospitalization, respiratory distress syndrome, multiple organ failure, pulmonary heart disease, and pneumothorax. The definitions and inclusion criteria for severe pulmonary TB in the included studies were extracted. Results: A total of 19 981 studies were identified and 100 studies were finally included, involving 8 309 patients with severe pulmonary TB. A total of 8 (8.00%) studies explicitly mentioned the definition of severe pulmonary TB, and 53 (53.00%) studies clearly defined the inclusion criteria for patients with severe pulmonary TB. A total of 5 definitions and 30 inclusion criteria were extracted. A total of 132 dichotomous variables and 113 continuous variables were included in the outcome indicators related to patients with severe pulmonary TB of concern in the studies. Conclusions: The definition and diagnostic criteria for severe TB are unclear, and there is an urgent need to develop a clear definition and diagnostic criteria to guide clinical practice.

[Analysis of the international application of healthy life expectancy].

Healthy life expectancy is based on life expectancy to further identify the healthy component, moving the assessment threshold from the mortality to the whole life cycle, receiving more and more attention worldwide. Nowadays, it has become one of the core indicators of national major strategy and plan. As a comprehensive indicator of health measurement, healthy life expectancy is complicated and multi-dimensional. Different social and cultural backgrounds have different understandings of health and choose different measurement dimensions. Overall, although high-income countries have different choices in their national health plan, healthy life expectancy without activity restriction is by far the most widely used indicator. This paper reviewed the concept and application of healthy life expectancy systematically, drawing on international practical experience to provide reference for the establishment of a healthy life expectancy indicator system in line with the Chinese national conditions.

[Prediction, analysis and application of learning curve of tooth preparation for all ceramic crowns of maxillary central incisors].

OBJECTIVE: To predict the learning curve of tooth preparation for all ceramic crowns of maxillary central incisors on phantom head simulators for graduate students participating in standardized dental resident training based on the modified Wright learning curve model, then to analyze and applicate the learning curve. METHODS: Twelve graduate students participating in standardized dental resident training were selected to prepare the resin maxillary central incisors on phantom head simulators for all ceramic crowns 4 times. The results of preparation were evaluated by 3 prosthetic experts with at least 10 years' experience focusing on the reduction, contour, taper, shoulder, finish line, margin placement, adjacent tooth injury, and preparation time for tooth preparation. The learning rate of tooth preparation was calculated by scores of tooth preparation of 4 times. The learning curve of tooth preparation was predicted based on the modified Wright learning curve model. According to the criteria of standardized training skill examinations for dental residents in Beijing, 80 was taken as the qualified standard score. The minimum training times for tooth preparation to satisfy the qualified standard score (80) was calculated, to analyze the characteristics of learning curve and evaluate the effectiveness of tooth preparation. RESULTS: The scores of 4 tooth preparation were 64.03±7.80, 71.40±6.13, 74.33±5.96, and 75.98±4.52, respectively. The learning rate was (106±4)%, which showed the learning curve an upward trend. There were no significant differences between the qualified standard score and the predicted scores of tooth preparation from the 5th preparation to the 13th preparation (P > 0.05). The predicted score of the 14th preparation was higher than the qualified standard score (P < 0.05). CONCLUSION: The trend of the learning curve of tooth preparation for all ceramic crowns of maxillary central incisors on phantom head simulators for graduate students participating in standardized dental resident training is upward, which predicts the minimum training times higher than the qualified standard score is 14 times.

Association between Preexisting Sarcopenia and Stroke in Patients with Type 2 Diabetes Mellitus.

OBJECTIVES: This propensity score-matched population-based cohort study compared stroke risk between patients with type 2 diabetes mellitus with and without preexisting sarcopenia. RESEARCH DESIGN AND METHODS: We used data from Taiwan's National Health Insurance Research Database for the period from January 2008 to December 2019. We recruited patients with type 2 diabetes mellitus and categorized them into two groups at a ratio of 1:1 on the basis of diagnosed sarcopenia. The matching variables were age, sex, income level, urbanization level, diabetes severity (adapted Diabetes Complications Severity Index [aDCSI Scores]), Charlson Comorbidity Index (CCI), other comorbidities associated with stroke, smoking status, medication use, and types of antidiabetic medications. The matching process yielded a final cohort of 104,120 patients (52,060 and 52,060 in the sarcopenia and nonsarcopenia groups, respectively) who were eligible for inclusion in subsequent analyses. RESULTS: In the multivariate Cox regression analysis, the adjusted hazard ratio (aHR; 95% CI) of stroke for the sarcopenia diabetes group compared with the control group was 1.13 (1.10, 1.16; P < 0.001), after controlling for age, sex, CCI, and aDCSI scores. The incidence rates of stroke for the sarcopenia and nonsarcopenia groups were 295.98 and 260.68 per 10,000 person-years, respectively. The significant IRR (95% CI) of stroke was 1.14 (1.09, 1.17) for the sarcopenia diabetes group compared with the nonsarcopenic diabetes group. CONCLUSION: Preexisting sarcopenia increased the risk of stroke in patients with type 2 diabetes mellitus.

Saturn Anomalous Myriametric Radiation, a New Type of Saturn Radio Emission Revealed by Cassini.

A new radio component namely Saturn Anomalous Myriametric Radiation (SAM) is reported. A total of 193 SAM events have been identified by using all the Cassini Saturn orbital data. SAM emissions are L-O mode radio emission and occasionally accompanied by a first harmonic in R-X mode. SAM's intensities decrease with increasing distance from Saturn, suggesting a source near Saturn. SAM has a typical central frequency near 13 kHz, a bandwidth greater than 8 kHz and usually drifts in frequency over time. SAM's duration can extend to near 11 hr and even longer. These features distinguish SAM from the regular narrowband emissions observed in the nearby frequency range, hence the name anomalous. The high occurrence rate of SAM after low frequency extensions of Saturn Kilometric Radiation and the SAM cases observed during compressions of Saturn's magnetosphere suggest a special connection to solar wind dynamics and magnetospheric conditions at Saturn.

Reflection and Refraction of the L-O Mode 5 kHz Saturn Narrowband Emission by the Magnetosheath.

The reflection-by-sheath mechanism of 5 kHz narrowband emissions (NB) at Saturn is confirmed by Cassini observations during several crossings of the magnetopause, which show that the 5 kHz NB can be prevented from escaping Saturn's magnetosphere. The L-O mode 5 kHz NB remained visible in areas of low plasma density but disappeared in regions of high plasma density. In three cases, NB disappeared immediately after the crossings of Saturn's magnetopause. A possible reflected NB event observed near the magnetosheath is discussed. This mechanism can help explain the 5 kHz NB observed at low latitudes outside the Enceladus plasma torus and their upper frequency limit variations. This mechanism significantly improves the current understanding of the 5 kHz NB.

[Benefits and risks of prolonged dual antiplatelet therapy after percutaneous coronary intervention with drug-eluting stent in patients with stable coronary artery disease and diabetes].

Objective: To compare the efficacy and safety of prolonged dual antiplatelet therapy (DAPT>1 year) in patients with stable coronary artery disease (CAD) and diabetes who were event-free at 1 year after percutaneous coronary intervention (PCI) with drug-eluting stent (DES) in a large and contemporary PCI registry. Methods: A total of 1 661 eligible patients were selected from the Fuwai PCI Registry, of which 1 193 received DAPT>1 year and 468 received DAPT ≤1 year. The primary endpoint was major adverse cardiac and cerebrovascular event (MACCE) and Bleeding Academic Research Consortium (BARC) type 2, 3 or 5 bleeding, MACCE was defined as a composite of all-cause death, myocardial infarction or stroke. Multivariate Cox regression analysis and inverse probability of treatment weighting (IPTW) Cox regression analysis were performed. Results: After a median follow-up of 2.5 years, patients who received DAPT>1 year were associated with lower risks of MACCE (1.4% vs. 3.2%; hazard ratio (HR) 0.412, 95% confidence interval (CI) 0.205-0.827) compared with DAPT ≤1 year, which was primarily caused by the lower all-cause mortality (0.1% vs. 2.6%; HR 0.031, 95%CI 0.004-0.236). Risks of cardiac death (0.1% vs. 1.5%; HR 0.051, 95%CI 0.006-0.416) and definite/probable ST (0.3% vs. 1.1%; HR 0.218, 95%CI 0.052-0.917) were also lower in patients received DAPT>1 year than those received DAPT ≤ 1 year. No difference was found between the two groups in terms of BARC type 2, 3, or 5 bleeding (5.3% vs. 4.1%; HR 1.088, 95%CI 0.650-1.821). Conclusions: In patients with stable CAD and diabetes who were event-free at 1 year after PCI with DES, prolonged DAPT (>1 year) provides a substantial reduction in ischemic cardiovascular events, including MACCE, all-cause mortality, cardiac mortality, and definite/probable ST, without increasing the clinically relevant bleeding risk compared with ≤ 1-year DAPT. Further well-designed, large-scale randomized trials are needed to verify the beneficial effect of prolonged DAPT in this population.

MicroRNA-543 inhibits the proliferation, migration, invasion, and epithelial-mesenchymal transition of triple-negative breast cancer cells via down-regulation of ACTL6A gene

PurposeTo investigate the effect of microRNA-543 (miR-543) on the proliferation, migration, invasion, and epithelial-mesenchymal transition (EMT) of triple-negative breast cancer (TNBC) cells, and the associated mechanism.MethodsHuman breast cancer cells (MDA-MB-231, HCC1937, and MCF-7, ZR-75–1) and normal human breast epithelial cell line (MCF10A) were transfected with miR-543 mimics or inhibitor using lipofectamine 2000. Quantitative reverse transcription polymerase chain reaction (qRT-PCR) and Western blotting were used to determine the mRNA and protein expression levels of miR-543, actin-like protein 6A (ACTL6A), vimentin, Snail, and E-cadherin in breast cancer cells/tissue. Cell counting kit-8 (CCK-8), wound-healing, and Transwell assays were used to measure the effect of miR-543 on TNBC cell proliferation, invasion, and migration. Overall survival was determined using data from Gene Expression Omnibus (GEO) and Cancer Genome Atlas (TCGA) databases. Bioinformatics analysis and luciferase reporter gene assay were used to determine the regulatory effect of miR-543 on ACTL6A.ResultsThe level of expression of miR-543 was significantly lower in breast cancer cells/tissue than in normal human breast epithelial cell/tissue (p < 0.05). MicroRNA-543 expression level was significantly reduced in TNBC cells/tissue, relative to the other breast cancer cells/normal breast tissue (p < 0.05). MicroRNA-543 significantly suppressed tumor growth and the proliferation, migration, invasion, and epithelial–mesenchymal transition (EMT) of TNBC cells, in mouse xenograft model (p < 0.05).

MicroRNA-543 inhibits the proliferation, migration, invasion, and epithelial-mesenchymal transition of triple-negative breast cancer cells via down-regulation of ACTL6A gene.

PURPOSE: To investigate the effect of microRNA-543 (miR-543) on the proliferation, migration, invasion, and epithelial-mesenchymal transition (EMT) of triple-negative breast cancer (TNBC) cells, and the associated mechanism. METHODS: Human breast cancer cells (MDA-MB-231, HCC1937, and MCF-7, ZR-75-1) and normal human breast epithelial cell line (MCF10A) were transfected with miR-543 mimics or inhibitor using lipofectamine 2000. Quantitative reverse transcription polymerase chain reaction (qRT-PCR) and Western blotting were used to determine the mRNA and protein expression levels of miR-543, actin-like protein 6A (ACTL6A), vimentin, Snail, and E-cadherin in breast cancer cells/tissue. Cell counting kit-8 (CCK-8), wound-healing, and Transwell assays were used to measure the effect of miR-543 on TNBC cell proliferation, invasion, and migration. Overall survival was determined using data from Gene Expression Omnibus (GEO) and Cancer Genome Atlas (TCGA) databases. Bioinformatics analysis and luciferase reporter gene assay were used to determine the regulatory effect of miR-543 on ACTL6A. RESULTS: The level of expression of miR-543 was significantly lower in breast cancer cells/tissue than in normal human breast epithelial cell/tissue (p < 0.05). MicroRNA-543 expression level was significantly reduced in TNBC cells/tissue, relative to the other breast cancer cells/normal breast tissue (p < 0.05). MicroRNA-543 significantly suppressed tumor growth and the proliferation, migration, invasion, and epithelial-mesenchymal transition (EMT) of TNBC cells, in mouse xenograft model (p < 0.05). CONCLUSIONS: miR-543 influences the biological behavior of TNBC cells by directly targeting ACTL6A gene. miR-543 could serve as a novel diagnostic and therapeutic target for TNBC.

[Clinical applicational comparison of digital impression and silicon rubber impression technique in posterior implant-supported single crown restoration].

Objective: To investigate the clinical effect and satisfaction of partially edentulous patients restored with posterior implant-supported single crown by digital impression technique and traditional silicon rubber impression, in order to provide clinical reference. Methods: Sixty-four partially edentulous patients who visited the Department of Prosthodontics, Guanghua School of Stomatology, Hospital of Stomatology, Sun Yat-sen University between March 2018 and January 2021 were enrolled. There were 31 male and 33 female, and the age of the group of patients was (49.3±13.3) years. The 113 implants placed in this group of patients were divided into digital impression group (n=70) and silicone rubber impression group (n=43) according to different impression techniques, and were restored with screw-retained full zirconia single crown, and the patients were followed up for 3-36 months after treatment. The implant survival rate, upper prosthesis, peri-implant soft tissue status, marginal bone loss and patients satisfaction were recorded to evaluate the clinical effects of two impression techniques in posterior implant-supported single crown. Results: In sixty-four patients, digital impression accounted for 62% (40/64), and silicon rubber impression accounted for 38% (24/64), the survival rate of 113 implants was 100% (113/113). The prevalence of interproximal contact loss, food impaction and mechanical complication of the upper full zirconia single crown restorations were 22.7% (41/181), 8.0% (9/113) and 2.7% (3/113) separately. All the mechanical complications were abutment screw loosening. All patients maintained good oral hygiene status, the incidences of peri-implantitis and peri-implant mucositis were 0.9% (1/113) and 4.4% (5/113) respectively. The marginal bone loss was (0.24±0.11) mm. The median of satisfaction visual analogue scale score in patients was 9-10, but no statistically significant differences of all the above results were observed between the two impression techniques (P>0.05). Conclusions: The clinical effect of partially edentulous patients restored with posterior implant-supported single crown using two different impression techniques is good and the satisfaction of patients is high, but the long-term clinical effect remains to be further observed.

Photoluminescence and magnetism integrated multifunctional black phosphorus probes through controllable PO bond orbital hybridization.

Biological probes with integrated photoluminescence and magnetism characteristics play a critical role in modern clinical diagnosis and surgical protocols combining fluorescence optical imaging (FOI) with magnetic resonance imaging (MRI) technology. However, traditional magnetic semiconductors can easily generate a spin splitting at the Fermi level and half-metallic electronic occupation, which will sharply reduce the radiation recombination efficiency of photogenerated carriers. To overcome this intrinsic contradiction, we propose a controllable oxidation strategy to introduce some particular PO bonds into black phosphorus nanosheets, in which the p orbital hybridization between P and O atoms not only provides some carrier recombination centers but also leads to a room-temperature spin polarization. As a result, the coexistence of photoluminescence and magnetism is realized in multifunctional black phosphorus probes with excellent biocompatibility. This work provides a new insight into integrating photoluminescence and magnetism together by intriguing atomic orbital hybridization.

Sirt7 protects chondrocytes degeneration in osteoarthritis via autophagy activation.

OBJECTIVE: Osteoarthritis (OA) is associated with decreased autophagy activity and imbalance of cell homeostasis in chondrocytes (CHs). Sirtuin 7 (Sirt7) is claimed to have the ability to activate the autophagy response. The aim of this study was to explore the function of Sirt7 in the development of OA involving autophagy by culturing human chondrocytes (CHs). PATIENTS AND METHODS: We collected knee joint cartilage from patients undergoing traumatic amputation and arthroscopic knee replacement. Protein and mRNA levels of collagen II, Sirt7, ULK1, Lc3, and Beclin1 were analyzed by Western blot and RT-PCR. CHs were isolated from the traumatic cartilage as a control group, and IL-1ß was used to induce CHs degeneration. The expression of Sirt7 gene was silenced by siRNA and upregulated by recombinant human Sirt7 protein (rh-Sirt7). An autophagy activator Tat-beclin 1 (Tat) was used to activate autophagy in cultural CHs. Expression of collagen II, Sirt7, ULK1, Lc3, and Beclin1 was determined by immunofluorescence, Western blot, and RT-PCR, respectively. RESULTS: The protein and mRNA levels of Collagen II, Sirt7, ULK1, Lc3-II, and Beclin1 expressions were decreased in OA cartilage compared with those in healthy cartilage. IL-1ß degenerated the CHs resulting in a reduction of collagen II, which also downregulated Sirt7, ULK1, Lc3-II, and Beclin1. Sirt7 deficiency accelerated the catabolism of collagen II and weakened the expression of ULK1, Lc3-II, and Beclin1. On the contrary, exogenous rh-Sirt7 played a positive role in these gene expressions. Finally, Tat alleviated the CHs degeneration by upregulating collagen II and activating ULK1, Lc3-II, and Beclin1, but incapable to mediate Sirt7 expression. CONCLUSIONS: Overall, these findings suggested that Sirt7 was suppressed in the degenerated cartilage. Sirt7 deficiency does harm to the autophagy level, affecting CHs metabolism, while the upregulation of Sirt7 activated autophagy and protected CHs degeneration. An appropriate increase in autophagy can protect CHs but has no effect on Sirt7 expression.

The Analysis of Drug-Resistant Gene Mutations of Mycobacterium tuberculosis by GeneChip in Lianyungang, China.

BACKGROUND: Tuberculosis (TB) has raised major global health concerns, especially for that caused by drug-resistant Mycobacterium tuberculosis (M. tuberculosis). The control of TB was hampered by time-consuming and insensitive diagnostic methods. GeneChip analysis is a rapid method for screening and identifying the gene mutations of M. tuberculosis. However, there was little relevant information about GeneChip analysis of M. tuberculosis in China. METHODS: To compare the performance of GeneChip analysis in the diagnosis of drug-resistant M. tuberculosis with traditional drug susceptibility testing (DST), 1,747 sputum specimens from 2014 to 2016 in Lianyungang of China were retrospectively analyzed. RESULTS: GeneChip analysis showed that the gene mutation site of M. tuberculosis to RFP resistance was 46.37% in rpoB 531 (TCG→TTG), and INH resistance was 69.89% in katG 315 (AGC→ACC). There was not significant different between GeneChip analysis and DST in detecting the resistance of M. tuberculosis to RPF or INH. CONCLUSIONS: GeneChip analysis could be regarded as a rapid and recommended method for early screening and identifying the drug resistance of M. tuberculosis.

[Authorization, translation, back translation and language modification of the Chinese version of the obturator functioning scale].

Objective: To cross-cultural adapt the English version of obturator functioning scale (OFS) to form a simplified Chinese version, to preliminarily verify its reliability and validity in clinic, and to provide an effective tool for evaluating the oral function and quality of life of patients with palatal defect and restored with obturators in China. Methods: The English version of the OFS was taken for forward translation, synthesis, back-translation, and reviewed by expert committee to develop a pre-testing simplified Chinese version. This scale contained demographic data, basic information of diseases, eating problems dimensions (3 items), speech problems dimensions (5 items), and other problems dimensions (7 items). From December, 2016 to December, 2018, forty-two patients who were treated in the Department of Prosthodontics, Guanghua School of Stomatology, Hospital of Stomatology, Sun Yat-sen University with palatal defect and restored with obturators were evaluated with OFS. Among them, there were 26 males, and 21-84 years old, and 16 females, who were 24-80 years old.The reliability and validity of the data were examined and analyzed. Results: The results showed that Cronbach's α coefficients of the overall scale and the three dimensions (eating problems, speech problems, and other problems) were 0.926, 0.938, 0.930, and 0.935, respectively. The internal consistency of the questionnaire was very good. The Spearman coefficients between each single dimension and the total score were 0.677, 0.792, and 0.860, respectively, suggesting that the scale convergence was good. The content validity index of 15 items was 0.905, indicating that the content validity was very good. Conclusions: The Chinese version of the OFS is exhibiting high reliability and validity, providing an effective evaluation tool of oral function and quality of life for Chinese patients with obturator prostheses to restore palate defects.

IL-9 exacerbates the development of chronic obstructive pulmonary disease through oxidative stress.

OBJECTIVE: To investigate the role of IL-9 in chronic obstructive pulmonary disease (COPD), and to explore its potential mechanism. MATERIALS AND METHODS: A mouse COPD model was established by exposure to cigarette smoke. COPD mice were then randomly assigned into two groups, including: the PBS group and the IL-9 antibody group. The above two groups were treated with phosphate-buffered saline (PBS) or IL-9 injection, respectively. The histopathological changes in lung tissues of mice were observed by hematoxylin-eosin (H&E) staining. Immunohistochemistry was performed to detect IL-9-positive (IL-9+) cells in lung tissues. Expression levels of IL-9, sIL-9R, STAT3, and p-STAT3 in peripheral blood of mice were determined by quantitative Real time-polymerase chain reaction (qRT-PCR), enzyme-linked immunosorbent assay (ELISA), and Western blot, respectively. In addition, the expression levels of superoxide dismutase (SOD), malondialdehyde (MDA), and reactive oxygen species (ROS) were detected. RESULTS: H&E staining results showed that the airway wall structure of COPD mice in the PBS group was irregular. Ciliated columnar epithelium exhibited marked degeneration, necrosis and shedding. Besides, numerous inflammatory cell infiltration, narrowing and rupture of the alveolar septa, and larger cysts fused by adjacent alveoli were observed. H&E staining also indicated that the structure of alveolar epithelium was severely impaired in COPD mice. However, the pathological changes in lung tissues of mice in the IL-9 antibody group were much milder than those of the PBS group. Immunohistochemistry results showed a significant deposition of IL-9+ cells in the lung tissues of the PBS group. Meanwhile, the mRNA and protein levels of IL-9, sIL-9R, and p-STAT3 in the PBS group were also remarkably higher than those of the IL-9 antibody group. In addition, SOD content in the PBS group was significantly decreased, whereas the levels of MDA and ROS were significantly increased than those of the IL-9 antibody group. CONCLUSIONS: IL-9 activated STAT3 and aggravated lung injury in COPD mice by increasing inflammatory and oxidative stress.

The therapeutic efficacy of pediatric ALL patients with MLL gene rearrangement treated with CCLG-ALL2008 protocol.

OBJECTIVE: In this study, we retrospectively evaluated the therapeutic efficacy of China Children Leukemia Group-ALL2008 (CCLG-ALL 2008) protocol in pediatric patients with mixed-lineage leukaemia (MLL) gene rearrangement of acute lymphoblastic leukemia (ALL) to identify the prognostic factors. PATIENTS AND METHODS: Six hundred and thirty-four patients with ALL were enrolled in this study between June 2008 and Dec 2014. High-risk group (HR) consisted of 217 cases, of which 28 cases were MLL related positive (first group), 22 cases were BCR/ABL positive (second group), and 167 cases were negative with MLL related or BCR/ABL (third group). The therapeutic efficacy was evaluated at the time points of day 8 (TP1), day 15 (TP2), day 33 (TP3) and 12th week (TP4) with the protocol, respectively. Overall-survival (OS) and relapse-free-survival (RFS) and treatment-related mortality (TRD) were analyzed as well. RESULTS: The first group accounted for 4.4% of all patients. Compared with the second and third group, the first group had more cases younger than 2 years, with initial leukocytes ≥50×109/L, and poor response on TP2. Moreover, patients older than 2 years old had a good 5 years OS (84% ± 9% vs. 37% ± 20%, p<0.05) and RFS (84% ± 9% vs. 29% ± 17%, p<0.05). There were no significant differences in the recurrence rate, TRD, 5 years OS and RFS among three groups. For the first group, compared with good response to prednisone, patients with poor response to prednisone had a poor 5 years RFS (41% ± 17% vs. 81% ± 10%, p<0.05). Multivariate Cox regression analysis identified that RFS and OS were influenced by such factors as age, MLL fusion partners, and prednisone response (p<0.05). CONCLUSIONS: Such factors as younger age than 2 years old, MLL/AF4 fusion gene, poor response to prednisone, or no complete remission (CR) on TP3 were poor prognostic parameters in predicting the outcome in childhood ALL with MLL gene rearrangement treated with CCLG-ALL 2008 protocol.

The Diagnostic Performance of Urine Flow Cytometer UF1000i for Urinary Tract Infections.

BACKGROUND: Urinary tract infections (UTIs) are one of the most common infectious diseases in clinic. Urine flow cytometry is receiving more and more attention due to its rapid forecast of UTIs. METHODS: The Urine Flow Cytometer UF1000i has a series of software programs to quantify bacteria (BACT) and white blood cells (WBC), and describe the scatter diagram of bacteria. The UTIs were predicted based on the cutoff values with the Receiver Operating Characteristic (ROC) curves of BACT and WBC counts. To evaluate the diagnostic performance of UF1000i for UTIs, the sensitivity and specificity of 889 urine samples were determined in comparison to the results of urine culture. Meanwhile the bacterial morphology indication of the UF1000i was evaluated in order to help doctors choose antibiotics. The angle of the scatter cloud with the x-axis was used to classify the infected bacteria as bacilli (< 30°) or cocci (≥ 30°). RESULTS: The best cutoff value of BACT counts for predicting UTIs was 119 per µL, and the sensitivity and specificity were 95.5% and 88.7%, respectively. While the best cutoff value of WBC counts was 81.5 per µL, and the sensitivity and specificity were 77.6% and 76.7%, respectively. In addition, the best cutoff values for females were 583 BACT per µL and 137.5 WBC per µL. They were much higher than for males (118 BACT per µL and 91 WBC per µL). The coincidence of the bacterial morphology information between the UF1000i software indication and the bacterial actual morphology identified by urine culture was 83% (bacilli) and 68% (cocci), respectively. CONCLUSIONS: Data demonstrated that the performance of BACT counts for UTIs is superior to WBC counts. In addition, the bacterial morphology could preliminarily be predicated by the scatter diagram. Since the urine flow cytometer UF1000i can provide the data of both BACT counts and the scatter diagram, the urine flow cytometry was regarded as a suitable method for screening UTIs. Moreover, it would be better to take gender into consideration when setting the best cutoff value for diagnosis of UTIs in clinic.

Suppressed CCL2 expression inhibits the proliferation of leukemia cells via the cell cycle protein Cyclin D1: preliminary in vitro data.

OBJECTIVE: Chemokine (C-C motif) ligand 2 (CCL2) is a member of the CC subfamily, which displays chemotactic activity for monocytes and basophils. This molecule plays a very important role in many solid tumors and shows changes in the bone marrow microenvironment. However, its role in acute myeloid leukaemia (AML) is still unclear. MATERIALS AND METHODS: In this study, we established a HL-60 cell line with CCL2 knockdown to explore its effect on leukemogenesis. Lentivirus with CCL2-knockdown was successfully constructed after screening effective CCL2 short hairpin RNA (shRNA) sequences and was transfected into HL-60 cells, which was further validated at the mRNA and protein levels by real-time polymerase chain reaction (PCR) and Western blotting, respectively. RESULTS: Low expression of CCL2 significantly decreased HL-60 cell growth by increasing the cell arrest at G1 phase by 12% more than controls. We applied RNA sequencing technology to discriminate the gene expression profiles between the cells with CCL2 knockdown and the controls, and Cyclin D1 was selected for further experiments as its expression level was significantly downregulated, which was validated at the mRNA and protein levels. Cyclin D1 knockdown experiments showed that the cell proliferation rate was evidently decelerated, and cell cycle analysis also indicated a similar pattern for CCL2. CONCLUSIONS: Our study revealed that Cyclin D1 is an effector that mediates CCL2's function in cell proliferation by blocking cells at G1 phase.