Clinical research on the relationship between the curve of Wilson and temporomandibular joint disorders.

OBJECTIVE: The aim of this study was to examine the relationship between the curve of Wilson (COW) and temporomandibular joint disorder (TMD). METHODS: The study cohort comprised patients aged 19-55 with malocclusion treated at our institution from January to July 2021. They were divided into a malocclusion with TMD group (TMD group) and a malocclusion without TMD group (non-TMB group) based on the diagnostic criteria of TMD. The study outcome was the differences in COW, measured via cone beam computed tomography (CBCT), and statistical analysis was performed using one-way analysis of variance and t-test. RESULTS: A total of 250 adult individuals were enrolled, including 162 females (age: 36.43 ± 11.00 years) and 88 males (age: 36.33 ± 9.88 years). Compared with the non-TMB group (n = 125), the TMD group (n = 125) had a significantly greater angle of COW (first molars: P = 0.002; second molars: P < 0.001), higher buccal inclination angle of molars in those with same side temporomandibular joint (TMJ) sounds than those with TMJ sounds (first molar: P = 0.000; second molar: P = 0.006) and greater the side with TMJ sounds (first molar: P < 0.001; second molar: P = 0.016). However, no difference was observed in the buccolingual axial inclination angle of molars between patients with and without TMJ sounds. CONCLUSION: The study reported the differences in malocclusion patients with and without TMB, which could be used as a reference by dentists to improve the treatment outcomes of these patients.

Thymoquinone induces apoptosis in temozolomide-resistant glioblastoma cells via the p38 mitogen-activated protein kinase signaling pathway.

Temozolomide (TMZ) can cross the blood-brain barrier (BBB) and deliver methyl groups to the purine (guanine) bases of DNA, leading to mispairing during DNA replication and subsequent cell death. However, increased expression of the repair enzyme methyl guanine methyltransferase (MGMT), which removes methyl groups from purine bases, counteracts methylation by TMZ. We evaluated the anticancer potential of thymoquinone (TQ), a hydrophobic flavonoid that inhibits resistance and induces apoptosis in various cancer cells, both in vitro and in vivo. In vitro experiments showed that compared with the Hs683 and M059J cell lines, U251 cells were more sensitive to TMZ. Compared to U251 cells, U251R cells, a TMZ drug-resistant strain established in this study, are characterized by increased expression of phosphorylated extracellular signal-regulated kinase (p-ERK) and MGMT. TQ treatments induced apoptosis in all cell lines. The p38 mitogen-activated protein kinase signal pathway was mainly activated in U251 and U251R cells; however, p-ERK and MGMT upregulation could not suppress TQ effects. Furthermore, si-p38 pretreatment of U251R cells in TQ treatments inhibited cell apoptosis. We speculate that TQ contributed to the phosphorylation and activation of p38, but not of ERK-induced apoptosis (irrespective of TMZ resistance). In vivo, U251R-derived tumors subcutaneously inoculated in nude mice exhibited significant tumor volume reduction after TQ or TQ + TMZ cotreatments. High-performance liquid chromatography assay confirmed the presence of TQ in murine brain tissues. Our findings demonstrate that TQ can effectively cross the BBB and function alone or in combination with TMZ to treat glioblastoma.

Abnormal liver tests are not sufficient for diagnosis of hepatic graft-versus-host disease in critically ill patients.

Hepatic graft-versus-host disease (HGVHD) contributes significantly to morbidity and mortality after hematopoietic stem cell transplantation (HSCT). Clinical findings and liver biomarkers are neither sensitive nor specific. The relationship between clinical and histologic diagnoses of HGVHD was assessed premortem and at autopsy. Medical records from patients who underwent HSCT at the National Institutes of Health (NIH) Clinical Center between 2000 and 2012 and expired with autopsy were reviewed, and laboratory tests within 45 days of death were divided into 15-day periods. Clinical diagnosis of HGVHD was based on Keystone Criteria or NIH Consensus Criteria, histologic diagnosis based on bile duct injury without significant inflammation, and exclusion of other potential etiologies. We included 37 patients, 17 of whom had a cholestatic pattern of liver injury and two had a mixed pattern. Fifteen were clinically diagnosed with HGVHD, two showed HGVHD on autopsy, and 13 had histologic evidence of other processes but no HGVHD. Biopsy or clinical diagnosis of GVHD of other organs during life did not correlate with HGVHD on autopsy. The diagnostic accuracy of the current criteria was poor (κ = -0.20). A logistic regression model accounting for dynamic changes included peak bilirubin 15 days before death, and an increase from period -30 (days 30 to 16 before death) to period -15 (15 days before death) showed an area under the receiver operating characteristic curve of 0.77. Infection was the immediate cause of death in 68% of patients. In conclusion, liver biomarkers at baseline and GVHD elsewhere are poor predictors of HGVHD on autopsy, and current clinical diagnostic criteria have unsatisfactory performance. Peak bilirubin and cholestatic injury predicted HGVHD on autopsy. A predictive model was developed accounting for changes over time. Further validation is needed.

[Association of miR-146a rs2910164 G/C polymorphism with its abnormal expression and risk of gastric cancer].

OBJECTIVE: To assess the influence of rs2910164 G/C single nucleotide polymorphism (SNP) of the miR-146a gene on its expression and susceptibility to gastric cancer. METHODS: Fifty three gastric cancer patients and six gastric cancer cell lines were selected for determining the miR-146a expression by Taqman quantitative PCR. A model was constructed to assess the influence of miR-146a overexpression on the growth of AGS gastric cancer cells. A case-control study involving 417 gastric cancer patients and 420 cancer-free individuals was then conducted, and the allelic and genotypic frequencies of the rs2910164 G/C SNP were compared. The genotypes of all subjects were determined by using a Taqman allelic discrimination assay. A Taqman assay was also used to quantify mature and pri-miR-146a transcripts among 65 gastric cancer patients with known genotypes. RESULTS: The expression of miR-146a was down-regulated among the 53 gastric cancer patients and six gastric cancer cell lines. Over-expression of miR-146a has suppressed the growth of gastric cancer by inhibiting the G1/S-phase transition of AGS cells. The case-control study showed that subjects with GC/CC genotypes had significantly lower risk for gastric cancer compared with those with GG genotype. In addition, miR-146a G/C SNP has significantly increased the level of mature miR-146a in those with GC/CC genotype compared with GG genotype. CONCLUSION: Down-regulation of miR-146a may play an important role in the pathogenesis of gastric cancer. The rs2910164 polymorphism of the miR-146a gene may reduce the risk of gastric cancer by influencing the processing of mature miR-146a.

Association of pet-keeping in home with self-reported asthma and asthma-related symptoms in 11611 school children from China.

OBJECTIVE: We conducted a cross-sectional study to investigate the associations between domestic pets and respiratory health in children. METHODS: We randomly recruited 11,611 school children from Zhongshan, a southern city in China. Information about the respiratory symptoms and disease history of the recruited children, the status of domestic pets, and other related risk factors were collected from March to July 2016. RESULTS: We identified cat-keeping at home increases the risk of persistent cough (OR, 1.77; 95%CI, 1.03-3.05); poultry-keeping at home increases the risk of current asthma (OR, 3.87; 95%CI, 1.08-13.92) and allergic rhinitis (OR, 1.84; 95%CI, 1.01-3.37); sleeping with pets increases the risk of persistent phlegm (OR, 5.04; 95%CI, 1.05-24.28), doctor-diagnosed asthma (OR, 3.35; 95%CI, 1.31-8.57) and current asthma (OR, 4.94; 95%CI, 1.05-23.31) in children. CONCLUSIONS: Cat-keeping and molds on the wall of the house had the multiplicative and additive interaction in doctor-diagnosed asthma. In conclusion, pet-keeping increased the risk of respiratory symptoms in children.

F0F1 ATP synthase regulates extracellular calcium influx in human neutrophils by interacting with Cav2.3 and modulates neutrophil accumulation in the lipopolysaccharide-challenged lung.

BACKGROUND: Neutrophils form the first line of innate host defense against invading microorganisms. We previously showed that F0F1 ATP synthase (F-ATPase), which is widely known as mitochondrial respiratory chain complex V, is expressed in the plasma membrane of human neutrophils and is involved in regulating cell migration. Whether F-ATPase performs cellular functions through other pathways remains unknown. METHODS: Blue native polyacrylamide gel electrophoresis followed by nano-ESI-LC MS/MS identification and bioinformatic analysis were used to identify protein complexes containing F-ATPase. Then, the identified protein complexes containing F-ATPase were verified by immunoblotting, immunofluorescence colocalization, immunoprecipitation, real-time RT-PCR and agarose gel electrophoresis. Immunoblotting, flow cytometry and a LPS-induced mouse lung injury model were used to assess the effects of the F-ATPase-containing protein complex in vitro and in vivo. RESULTS: We found that the voltage-gated calcium channel (VGCC) α2δ-1 subunit is a binding partner of cell surface F-ATPase in human neutrophils. Further investigation found that the physical connection between the two proteins may exist between the F1 part (α and ß subunits) of F-ATPase and the α2 part of VGCC α2δ-1. Real-time RT-PCR and PCR analyses showed that Cav2.3 (R-type) is the primary type of VGCC expressed in human neutrophils. Research on the F-ATPase/Cav2.3 functional complex indicated that it can regulate extracellular Ca2+ influx, thereby modulating ERK1/2 phosphorylation and reactive oxygen species production, which are typical features of neutrophil activation. In addition, the inhibition of F-ATPase can reduce neutrophil accumulation in the lungs of mice that were intratracheally instilled with lipopolysaccharide, suggesting that the inhibition of F-ATPase may prevent neutrophilic inflammation-induced tissue damage. CONCLUSIONS: In this study, we identified a mechanism by which neutrophil activity is modulated, with simultaneous regulation of neutrophil-mediated pulmonary damage. These results show that surface F-ATPase of neutrophils is a potential innate immune therapeutic target.

LncRNA-DQ786227-mediated cell malignant transformation induced by benzo(a)pyrene.

It has recently been found that the new class of transcripts, long non-coding RNAs (lncRNAs), are pervasively transcribed in the genome. LncRNAs are a large family of non-coding RNAs and regulate many protein-coding genes. Growing evidence indicates that lncRNAs may play an important functional role in cancer biology. Emerging data have shown that lncRNAs are closely related to the occurrence and development of lung cancer. However, the function and mechanism of lncRNAs in lung cancer remain elusive. Here, we investigated the role of a novel lncRNA in transformed human bronchial epithelial cells induced by benzo(a)pyrene. After establishing the transformed cell model using the BEAS-2B cell line in vitro, we found that expression of lncRNA-DQ786227 was high and changed during the transformation of BEAS-2B cells. Silencing of lncRNA-DQ786227 expression in malignant transformed BEAS-2B cells led to inhibition of cell proliferation and colony formation, and increased apoptosis. LncRNA-DQ786227 dramatically promoted the ability of BEAS-2B-T cells to form colonies in vitro and develop tumors in nude mice. These findings revealed that lncRNA-DQ786227 acts as an oncogene in malignantly transformed BEAS-2B cells induced by benzo(a)pyrene. The identification of lncRNA could provide new insight into the molecular mechanisms of chemical carcinogenesis.

Kinetic mechanism of translocation and dNTP binding in individual DNA polymerase complexes.

Complexes formed between phi29 DNA polymerase (DNAP) and DNA fluctuate discretely between the pre-translocation and post-translocation states on the millisecond time scale. The translocation fluctuations can be observed in ionic current traces when individual complexes are captured atop the α-hemolysin nanopore in an electric field. The presence of complementary 2'-deoxynucleoside triphosphate (dNTP) shifts the equilibrium across the translocation step toward the post-translocation state. Here we have determined quantitatively the kinetic relationship between the phi29 DNAP translocation step and dNTP binding. We demonstrate that dNTP binds to phi29 DNAP-DNA complexes only after the transition from the pre-translocation state to the post-translocation state; dNTP binding rectifies the translocation but it does not directly drive the translocation. Based on the measured time traces of current amplitude, we developed a method for determining the forward and reverse translocation rates and the dNTP association and dissociation rates, individually at each dNTP concentration and each voltage. The translocation rates, and their response to force, match those determined for phi29 DNAP-DNA binary complexes and are unaffected by dNTP. The dNTP association and dissociation rates do not vary as a function of voltage, indicating that force does not distort the polymerase active site and that dNTP binding does not directly involve a displacement in the translocation direction. This combined experimental and theoretical approach and the results obtained provide a framework for separately evaluating the effects of biological variables on the translocation transitions and their effects on dNTP binding.

Dynamics of the translocation step measured in individual DNA polymerase complexes.

Complexes formed between the bacteriophage phi29 DNA polymerase (DNAP) and DNA fluctuate between the pre-translocation and post-translocation states on the millisecond time scale. These fluctuations can be directly observed with single-nucleotide precision in real-time ionic current traces when individual complexes are captured atop the α-hemolysin nanopore in an applied electric field. We recently quantified the equilibrium across the translocation step as a function of applied force (voltage), active-site proximal DNA sequences, and the binding of complementary dNTP. To gain insight into the mechanism of this step in the DNAP catalytic cycle, in this study, we have examined the stochastic dynamics of the translocation step. The survival probability of complexes in each of the two states decayed at a single exponential rate, indicating that the observed fluctuations are between two discrete states. We used a robust mathematical formulation based on the autocorrelation function to extract the forward and reverse rates of the transitions between the pre-translocation state and the post-translocation state from ionic current traces of captured phi29 DNAP-DNA binary complexes. We evaluated each transition rate as a function of applied voltage to examine the energy landscape of the phi29 DNAP translocation step. The analysis reveals that active-site proximal DNA sequences influence the depth of the pre-translocation and post-translocation state energy wells and affect the location of the transition state along the direction of the translocation.

Direct observation of translocation in individual DNA polymerase complexes.

Complexes of phi29 DNA polymerase and DNA fluctuate on the millisecond time scale between two ionic current amplitude states when captured atop the α-hemolysin nanopore in an applied field. The lower amplitude state is stabilized by complementary dNTP and thus corresponds to complexes in the post-translocation state. We have demonstrated that in the upper amplitude state, the DNA is displaced by a distance of one nucleotide from the post-translocation state. We propose that the upper amplitude state corresponds to complexes in the pre-translocation state. Force exerted on the template strand biases the complexes toward the pre-translocation state. Based on the results of voltage and dNTP titrations, we concluded through mathematical modeling that complementary dNTP binds only to the post-translocation state, and we estimated the binding affinity. The equilibrium between the two states is influenced by active site-proximal DNA sequences. Consistent with the assignment of the upper amplitude state as the pre-translocation state, a DNA substrate that favors the pre-translocation state in complexes on the nanopore is a superior substrate in bulk phase for pyrophosphorolysis. There is also a correlation between DNA sequences that bias complexes toward the pre-translocation state and the rate of exonucleolysis in bulk phase, suggesting that during DNA synthesis the pathway for transfer of the primer strand from the polymerase to exonuclease active site initiates in the pre-translocation state.

Synthesis and characterization of waterborne polyurethane/Cu(II)-loaded hydroxyapatite nanocomposites with antibacterial activity.

A novel kind of environmentally friendly nanocomposites, waterborne polyurethane (WBPU)/Cu(II)-loaded hydroxyapatite (CuHAp), with improved physical properties and antibacterial activity have been prepared via in-situ polymerization from functionalized CuHAp nanoparticles (CuHAp NPs). The interaction of the CuHAp NPs with isophorone diisocyanate to form the functionalized CuHAp NPs containing isocyanate groups (CuHAp-g-NCO) has been studied. The microstructure and particle distribution of the nanocomposites were observed using scanning electron microscopy. The improvements of mechanical properties, thermal stability and water resistance of the nanocomposites have also been evaluated. Finally, the antibacterial activity was tested against G(-) Escherichia coli and G(+) Staphylococcus aureus by the zone of inhibition test and the direct contact test. The long-lasting antibacterial activity was studied by measuring antibacterial ability of the nanocomposites after being immersed in water. The results indicate that WBPU incorporation with CuHAp NPs shows strong antibacterial activity upon contact, and long-lasting antibacterial property.