NDUFB11 and NDUFS3 regulate arterial atherosclerosis and venous thrombosis: Potential markers of atherosclerosis and venous thrombosis.

Atherosclerosis is a chronic disease that thickens the blood vessel walls and narrows the lumen. Venous thrombosis is a blood clot that forms in the body's deep veins or pulmonary arteries. However, the relationship between NDUFB11 and NDUFS3 and atherosclerosis and venous thrombosis is unclear. We employed data files that combined atherosclerosis and chronic stress groups. Subsequently, we conducted differential gene expression analysis (DEGs) and performed weighted gene co-expression network analysis (WGCNA). We constructed and analyzed a protein-protein interaction (PPI) network. Further analyses included functional enrichment analysis, gene set enrichment analysis (GSEA), gene expression heatmaps, immune infiltration analysis, and mRNA analysis. By comparing our findings with the Comparative Toxicogenomics Database (CTD), we identified the most relevant diseases associated with the core genes. Additionally, we utilized TargetScan to screen for miRNAs regulating the central DEGs. To validate our results, we conducted Western Blot experiments at the cellular level. A total of 1747 DEGs were co-identified. According to the Gene Ontology (GO) analysis of differentially expressed genes, they were primarily enriched in mitochondrial gene expression, mitochondrial envelope, organelle membrane, and mitochondrial inner membrane categories. Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis revealed that the target cells were mainly enriched in metabolic pathways, ribosomes, and histidine metabolism. The intersection of enriched terms from both GO and KEGG analyses showed significant enrichment in mitochondrial gene expression, mitochondrial envelope, organelle inner membrane, ribosomal structural constituents, histidine metabolism, and oxidative phosphorylation. Eight core genes were identified, including NDUFS5, UQCRQ, COX6C, COX7B, ATP5ME, NDUFS3, NDUFA3, and NDUFB11. The gene expression heatmap demonstrated that core genes (NDUFB11 and NDUFS3) were downregulated in atherosclerosis with venous thrombosis samples and upregulated in normal samples. CTD analysis revealed that the core genes NDUFB11 and NDUFS3 were associated with pain, arterial diseases, atherosclerosis, arteritis, venous thrombosis formation, and venous thromboembolism. We added Western Blot basic cell experiment for verification. NDUFB11 and NDUFS3 are downregulated in atherosclerosis and venous thrombosis, associated with poorer prognosis, and may serve as potential biomarkers for both diseases.

Inhibiting the transcription and replication of Ebola viruses by disrupting the nucleoprotein and VP30 protein interaction with small molecules.

Ebola virus (EBOV) causes hemorrhagic fever in humans with high morbidity and fatality. Although over 45 years have passed since the first EBOV outbreak, small molecule drugs are not yet available. Ebola viral protein VP30 is a unique RNA synthesis cofactor, and the VP30/NP interaction plays a critical role in initiating the transcription and propagation of EBOV. Here, we designed a high-throughput screening technique based on a competitive binding assay to bind VP30 between an NP-derived peptide and a chemical compound. By screening a library of 8004 compounds, we obtained two lead compounds, Embelin and Kobe2602. The binding of these compounds to the VP30-NP interface was validated by dose-dependent competitive binding assay, surface plasmon resonance, and thermal shift assay. Moreover, the compounds were confirmed to inhibit the transcription and replication of the Ebola genome by a minigenome assay. Similar results were obtained for their two respective analogs (8-gingerol and Kobe0065). Interestingly, these two structurally different molecules exhibit synergistic binding to the VP30/NP interface. The antiviral efficacy (EC50) increased from 1 µM by Kobe0065 alone to 351 nM when Kobe0065 and Embelin were combined in a 4:1 ratio. The synergistic anti-EBOV effect provides a strong incentive for further developing these lead compounds in future studies.

[Start-up Performance of Low-substrate Anaerobic Ammonium Oxidation Under Different COD Concentrations].

An anaerobic sequencing batch reactor(ASBR)was used to treat low-substrate simulated wastewater with NH4+-N and NO2--N concentrations of (25.00±0.40) mg·L-1 and (33.00±0.60) mg·L-1, respectively. The COD concentrations were controlled at 5.00, 15.00, 30.00, and 50.00 mg·L-1 by adding sodium acetate, and its effects on start-up of anaerobic ammonia oxidation (ANAMMOX) were investigated under the temperature of 30℃. The results showed that ① The start-up of ANAMMOX could be achieved successfully after 74, 94, 106, and 129 days. The nitrogen removal efficiency was optimum when the COD concentration was between 15.00 and 30.00 mg·L-1. In the steady phase, the average effluent concentrations of NH4+-N were 1.98 and 1.89 mg·L-1, the average effluent concentrations of NO2--N were below 0.62 mg·L-1, and the average effluent concentrations of TN were 2.37 and 2.28 mg·L-1. ② The average contribution of heterotrophic denitrification to nitrogen removal decreased to 4.78%, 9.59%, 10.21%, and 36.50%, respectively, during start-up process. The average contribution of ANAMMOX to nitrogen removal gradually increased to 95.22%, 90.41%, 89.79%, and 63.50%, respectively. ③ The activities of ANAMMOX exceeded denitrification activities at 44, 76, 86, and 114 days, respectively, which finally reached 0.700, 0.690, 0.670, and 0.510 mg·(g·h)-1, and the denitrification activities were 0.110, 0.130, 0.240 and 0.410 mg·(g·h)-1, respectively. Thus, the research results have provided references for the application of ANAMMOX to engineering.

G-quadruplex fluorescent probe-mediated real-time rolling circle amplification strategy for highly sensitive microRNA detection.

Real-time PCR has revolutionized PCR from qualitative to quantitative. As an isothermal DNA amplification technique, rolling circular amplification (RCA) has been demonstrated to be a versatile tool in many fields. Development of a simple, highly sensitive, and specific strategy for real-time monitoring of RCA will increase its usefulness in many fields. The strategy reported here utilized the specific fluorescence response of thioflavin T (ThT) to G-quadruplexes formed by RCA products. Such a real-time monitoring strategy works well in both traditional RCA with linear amplification efficiency and modified RCA proceeded in an exponential manner, and can be readily performed in commercially available real-time PCR instruments, thereby achieving high-throughput detection and making the proposed technique more suitable for biosensing applications. As examples, real-time RCA-based sensing platforms were designed and successfully used for quantitation of microRNA over broad linear ranges (8 orders of magnitude) with a detection limit of 4 aM (or 0.12 zmol). The feasibility of microRNA analysis in human lung cancer cells was also demonstrated. This work provides a new method for real-time monitoring of RCA by using unique nucleic acid secondary structures and their specific fluorescent probes. It has the potential to be extended to other isothermal single-stranded DNA amplification techniques.

[XANES study of lead speciation in duckweed].

Qixiashan lead-zinc mine of Nanjing was one of the largest lead zinc deposits in East China Its exploitation has been over 50 years, and the environmental pollution has also been increasing. The lead concentration in the local environment was high, but lead migration and toxic mechanism has not been clear. Therefore, biogeochemistry research of the lead zinc mine was carried out. Using ICP-MS and Pb-L III edge XANES, lead concentration and speciation were analyzed respectively, and duckweed which can tolerate and enriched heavy metals was found in the pollution area. The results showed that the lead concentration of duckweed was 39.4 mg x kg(-1). XANES analysis and linear combination fit indicated that lead stearate and lead sulfide accounted for 65% and 36.9% respectively in the lead speciation of duckweed, suggesting that the main lead speciation of duckweed was sulfur-containing lead-organic acid.

[Impact of magnetic field exposure on cardiac autonomic tone and inducibility of atrial fibrillation in dogs].

OBJECTIVE: To observe the maximal heart rate changes, atrioventricular (A-V) conduction block and atrial fibrillation (AF) inducibility in dogs with vagosympathetic trunk exposed to electromagnetic fields (EMFs). METHODS: The vagosympathetic trunk of adult dogs was separated and exposed to EMFs 0.043 kHz (2.87 microG, n = 5) and to EMFs 2 kHz (0.34 microG, n = 6) for two to three hours. Simultaneously, the vagosympathetic trunk was stimulated with 20 Hz frequency and 1 - 8 V intensity for 0.1 ms. Heart rate, presence of A-V conduction block and AF inducibility were determined. RESULTS: After 5-minutes exposure to EMFs 0.043 kHz (2.87 microG), the maximal heart rate decreased 29%, the voltage applied to vagosympathetic trunk required to induce A-V conduction block decreased by 60% in experimental group versus 5% increase in control group. This effect lasted 2 to 3 hours. While vagosympathetic trunk exposure to EMFs 2 kHz (0.34 microG) was associated with significant increase in the incidence of atrial premature beats, atrial tachycardia and AF, these effects could be blocked by propranolol and atropine. CONCLUSIONS: Our results showed that 0.043 kHz (2.87 microG) EMFs exposure might reduce while 2 kHz (0.34 microG) EMFs exposure might increase AF inducibility. Our study thus suggested autonomic nervous system of dogs could be affected by EMFs exposure and 0.043 kHz (2.87 microG) EMFs exposure might be a novel option for AF prevention.

[Therapeutic effect of endoscopic vidian neurectomy on moderate-severe persistent allergic rhinitis].

OBJECTIVE: To evaluate the efficacy of endoscopic vidian neurectomy in the management of moderate-severe persistent allergic rhinitis, and to explore its possible mechanism. METHODS: One hundred and ninety-one patients with moderate-severe persistent allergic rhinitis were divided into three groups: endoscopic vidian neurectomy was carried out in 71 patients (group A), partial inferior turbinectomy and/or septal-plasty in 39 patients (group B), and 81 patients were as control (group C). The life quality was assessed at 6 month, 1 year and 3 years after operation using rhinoconjunctivitis quality of life questionnaire (RQLQ) and visual analogue scale (VAS). RESULTS: Among 191 cases, one hundred and forty-five cases had complete follow-up documents. The average score of RQLQ and VAS score (x(-) ± s) were significantly decreased at the time of 6 months (0.84 ± 0.41, 2.55 ± 1.57), 1 year (0.91 ± 0.43, 2.63 ± 1.71) and 3 years (1.03 ± 0.46, 2.81 ± 1.75) after endoscopic vidian neurectomy than scores before operation (2.25 ± 0.49, 7.34 ± 1.11), F = 115.45, 133.09, respectively, P < 0.001, and also significantly lower than scores in patients in group B or control group at the same period after treatment. By patient's self-evaluation, the ratio of greatly-improved, improved and not-improved was respectively, 65.5% (38 cases), 24.1% (14 cases), 10.4% (6 cases), and significantly higher in patients in group A than in patients in group B (U = 237.0, P < 0.001) and group C (U = 246.0, P < 0.001). There was no severe complication in all patients observed. CONCLUSION: Endoscopic vidian neurectomy is an effective and safe technique in the management of moderate-severe persistent allergic rhinitis.

[Efficacy of sequential ablation of sinus atrial node fat pad and atrial ventricular node fat pad on inducibility of atrial fibrillation evoked by vagus trunk stimulation].

OBJECTIVE: To explore the efficacy of sequential ablation of epicardial fat pad on inducibility of atrial fibrillation (AF) evoked by stimulating vagus trunk. METHODS: Eighteen adult mongrel dogs were randomly divided into 2 groups (n = 9 each): Group A underwent pre-ablation of sinus-atrial node fad pad (SANFP) and subsequent ablation of atria-ventricular node fad pad (AVNFP). Group B underwent pre-ablation of AVNFP and subsequent ablation of SANFP. AF was induced by high-frequency electrical stimulation of bilateral vagus trunks. The AF inducibility and effective refractory period (ERP) changes during vagus trunk stimulation were examined before and after ablation in atria and pulmonary veins. RESULTS: (1) AF could be induced by vagus trunk stimulation and the incidence was higher during right vagus trunk (RVG) stimulation than left vagus trunk (LVG) stimulation [(60.0 ± 0.0)% vs (18.4 ± 22.1)%]. (2) SANFP ablation significantly attenuated AF inducibility with LVG stimulation and RVG stimulation at 2 V (decreased 67.0% and 72.0%, respectively). Subsequent AVNFP ablation after SANFP ablation further reduced AF inducibility with LVG and RVG stimulation at 2 V (decreased 100.0% and 95.5%, respectively). (3) AVNFP ablation (decreased 95.7% and 96.3%, respectively) and subsequent SANFP ablation after AVNFP ablation (decreased 98.0% and 100.0%, respectively) significantly attenuated AF inducibility with LVG stimulation and RVG stimulation at 2V. (4) Vagal stimulation induced ERP shortening was significantly attenuated by isolated SANFP ablation or AVNFP. Subsequent AVNFP ablation after SANFP induced significant ERP shortening in right atrial site compared with isolated SANFP ablation. However, changes of ERP shortening were similar between AVNFP ablation and subsequent SANFP ablation after AVNFP ablation. CONCLUSIONS: Epicardial fat pad ablation reduced the AF inducibility and prolonged ERP of atria and pulmonary veins during vagus trunk stimulation. AVNFP, as the "integration centers" modulating the vagal innervation to the atria, may be the more effective target of ablation for treating AF.

[Nerve remodeling in a canine model of atrial fibrillation induced by 48 hours right atrial pacing].

OBJECTIVE: To evaluate the nerve remodeling induced by 48 hours right atrial pacing in a canine model. METHODS: Rapid right atrial pacing (600 beats/min) was performed in 6 mongrel dogs of either sex for 48 hours to induce sustained atrial fibrillation (AF). Six dogs without pacing served as controls. Cardiac nerves were immunocytochemically stained using anti-growth-associated protein 43 (GAP43) and anti-choline acetyltransferase (CHAT) antibodies to compare nerve sprouting and pneumogastric nerve remodeling between the 2 groups. RESULTS: In dogs with AF, the GAP43-positive and CHAT-positive nerve densities in the left atrium, left auricular appendage, right atrium and right auricular appendage were significantly higher than in control animals (all P < 0.05). Moreover, nerve density was significantly higher in the right atrium than in the left atrium in dogs with AF. Microscopic examinations revealed an inhomogeneous distribution of cardiac nerves. CONCLUSION: Significant nerve sprouting and pneumogastric nerve remodeling were evidenced in the right and left atrium in a canine model of sustained AF induced by 48 hours right atrial pacing.

[Development of microsatellites of Dendrobium officinale and its application in purity identification of germplasm].

Simple sequence repeat (SSR) was used to investigate the genetic diversity and structure of Dendrobium officinale. A total of 15 primer pairs with stable and repeatable polymorphism were screened out from 60 SSR primer pairs developed by the method of microsatellite enrichment by magnetic beads. Forty-eight samples of Dendrobium officinale were analyzed in genetic polymorphism. These loci were polymorphic and displayed 3 to 9 alleles per locus with a mean number of 6.1. The observed and expected heterozygosities ranged from 0.60 to 0.85 and from 0.49 to 0.85 respectively. The polymorphic information content (PIC) of each SSR locus varied from 0.437 to 0.829 with an average of 0.702. Fifteen primer pairs were used in Dendrobium cross-species amplification and totally 13 primer pairs were proved to have the transferability in D. officinale related species. In addition, 500 tissue culture plantlets of D. officinale were tested for purity identification by means of PCR amplification with four SSR primer pairs. The results showed that SSR technique is a feasible, simple and inexpensive method for determining adulterants in germplasm identification.

[Experimental study on monitoring gene expression by noninvasive method in rabbit VX2 liver tumor model].

OBJECTIVE: To investigate the feasibility of monitoring therapeutic effect of adenovirus vector containing IL12-IRES-CKb gene on a rabbit VX2 liver tumor model by using phosphorous-31 magnetic resonance spectroscopy (31P MRS). METHODS: A total of 18 healthy New Zealand White rabbits were used to generate animal models by implanting VX2 tumor chips into livers through laparotomy. Tumor-bearing animals were randomly divided into three groups and were injected with AdCMVIL12-IRES-CKb, AdCMV-Empty and saline respectively via ear veins. 31P MRS scan was performed after animals were fed with creatine solution for five days. Animals were euthanized thereafter and tumors were removed for pathological examination, immunohistochemistry (IHC) staining and protein analysis (Western blot). RESULTS: The intrahepatic and seral expressions of creatine kinase (CKb) and IL-12 were detected only in AdCMVIL12-IRES-CKb group. Tumor diameters pre- and post- treatment in three groups were 1.63+/-0.04 vs 1.62+/-0.03 in AdCMVIL12-IRES-CKb group (P = 0.229), 1.59+/-0.05 vs 1.84+/-0.11 in AdCMV-Empty group (P = 0.003) and 1.60+/-0.02 vs 2.07+/-0.12 in saline group (P = 0.001), respectively. Pcr Changes between pre- and post- treatment among the three groups were compared (F = 6.235, P value is less than 0.05). PCr increased significantly in AdCMVIL12-IRES-CKb group as compared to AdCMV-Empty (P = 0.004) and saline group (P = 0.049), whereas no change found between AdCMV-Empty and saline group (P = 0.153). CONCLUSION: 31P MRS, an effective and non-invasive functional imaging method, can be used to monitor the therapeutic effect of adenovirus vector containing IL12-IRES-CKb gene on rabbit VX2 liver tumor model through detecting metabolic product of imaging reporter gene CKb (pCr).

[Role of folate receptor 1 in paclitaxel-resistance of nasopharyngeal carcinoma cells].

OBJECTIVE: To provide experimental evidence for the folate receptor 1 (FOLR1) mediated targeted cancer therapy and resistance reversal, the FOLR1 expression differences in nasopharyngeal carcinoma cells (CNE-1) and immortalized normal nasopharyngeal cells (NP69) and the correlation between FOLR1 expression and paclitaxel resistance index in nasopharyngeal carcinoma were investigated. METHODS: The expressions of FOLR1 in CNE-1, CNE-1/Taxol (paclitaxel-resistance cells) and NP69 was detected by cDNA microarray, reverse transcriptase-polymerase chain reaction (RT-PCR), Western blot and immunocytochemistry. Proliferation inhibition rates of CNE-1 and CNE-1/Taxol cells were measured by colony formation assay after treated by short interfering RNA (siRNA) of FOLR1. RESULTS: The expressions of FOLR1 gene in CNE-1/Taxol cells and CNE-1 cells were 2636.0 and 176.0, respectively. The expression of FOLR1 was not detected in the NP69 by semi-quantative RT-PCR and Western blot. The high expression of FOLR1 in CNE-1/Taxol was verified by semi-quantative RT-PCR, and its expression level was positively correlated to the degree of drug-resistance (r(2) = 0.8719). The results were also validated by Western blot and immunocytochemistry. The sensitivity of CNE-1/Taxol to paclitaxel significantly increased after inhibition of FOLR1 gene expression by siRNA, and its IC(50) value was decreased by 59.6% (t = 6.92, P < 0.01). CONCLUSIONS: The expression of FOLR1 is closely related to the occurrence of NPC and Taxol resistance. FOLR1 gene may be one of the important target molecules in NPC treatment and reversion of the paclitaxel-resistance in NPC.

[Analysis and authentication of cpDNA psbA-trnH regions of Dendrobium species of fengdous].

The psbA-trnH regions of Dendrobium species of Fengdous were sequenced by our research group. The psbA-trnH sequences of fifteen Dendrobium species were analyzed with software MEGA 4.0. The results showed that the lengths of sequences varied from 721 to 767 bp. The variable sites were 42 while the informative sites were 11. Genetic distances were calculated using the Kimura 2-parameter model. Genetic distances varied from 0.0013-0.0183 among fifteen species while the average genetic distance was 0.0148. The interspecies differences of psbA-trnH regions were demonstrated. Six indels happened in this fragment, which led to the great difference of sequence lengths among fifteen species. We found that there were no population differences in the psbA-trnH region of various species of Fengdous so far. By using the database of various Dendrobium species of Fengdous and two genetics software, the botanical origin of the inspected species of Fengdous was authenticated successfully by sequencing the psbA-trnH regions. The psbA-trnH region of cpDNA can be used as a candidate marker for authentication of Dendrobium species of Fengdous.

[Effect of seleno-arginine on cellular immunological function in D-gal aging mice].

AIM: To study the effect of seleno-argnine on cellular immunological function in D-gal aging mice. METHODS: The mice were divided into normal group, model group, low seleno-argnine group(L-SeArg) and high seleno-argnine group (H-SeArg). D-galactose induced aging model was set up by s.c. injection for 42 days, i.g. method was used to add Seleno-argnine. Spleen index(SI) and thymus index (TI) were measured. The ability of Spleen cell multiplication and NK cell killing activity were determined by MTT colorimetry. The changes of CD40 and CD40L expression were observed by immunofluorescent assay. RESULTS: Compared with the model mice, seleno-argnine increased spleen index and thynus index, enhanced the ability of spleen cell multiplication, enhanced the expression of CD40 and CD40L, and accelerated the NK cell killing activity. Between the two seleno-argnine groups, the effect of L-SeArg on cellular immunological function was more obvious in D-gal aging mice.The result were statistic ally significantly compared with the model mice(P<0.01). CONCLUSION: D-gal injection can decrease the cellular immunological function of mice. Seleno-argnine can activate immune cells, enhance the cellular immunological function of mice, reduce the toxic effect of D-gal and delay the aging process.

[Causes and revision endoscopic sinus surgery for recurrent sinusitis].

OBJECTIVE: To investigate the causes of revision endoscopic sinus surgery (RESS), and to evaluate the clinical effect and experience of RESS in the patients with recurrent sinusitis. METHODS: Before the revision surgery, 168 patients (225 sides) with recurrent sinusitis were examed by CT scans and nasal endoscopy. All patients were operated by revision endoscopic sinus surgery. RESULTS: Among the 168 patients, 95 cases (121 sides) had incomplete middle turbinate, 51 cases (75 sides) had maxillary sinus ostium stenosis, 85 cases (117 sides) had nasal cavity adhesion, 48 cases (57 sides) had uncinate process residual, 38 cases (65 sides) had ethmoid hyperostosis, and 25 cases had nasal septum deviation. One hundred and six patients (134 sides) were cured, 40 patients (56 sides) were improved, useful 22 patients (35 sides) were ineffective. CONCLUSION: The most common surgical causes of failures in endoscopic sinus surgery are the disease area remnant, nasal cavity adhesion, maxillary sinus ostium stenosis, uncinate process residual, ethmoid hyperostosis, and nasal septum deviation. Revision endoscopic sinus surgery is a useful way to treat recurrent sinusitis. Preoperative CT scan, correct choice of the anatomic marks of orientation and direction function are the key to operation successful.