[The analysis of a pedigree with hereditary coagulation factor â ¤ deficiency caused by compound heterozygous variation of F5 gene].

Objective: To analyze the gene variation of a genetic coagulation factor Ⅴ (FⅤ) deficiency pedigree and explore the molecular pathogenesis. Methods: The proband was a 32 years old female. The patient was prone to nose bleeding since childhood which was usually self-healed. On March 10, 2021, the proband went to the First Affiliated Hospital of Air Force Medical University for treatment of knee hematoma caused by a fall. None of the family members reported any history of bleeding. The prothrombin time (PT), activated partial thromboplastin time (APTT) and FⅤ activity (FⅤ: C) were detected by clotting method and the FⅤ antigen (FⅤ: Ag) was tested with enzyme-linked immunosorbent assay (ELISA). All exons and flanks of F5 gene were determined by Sanger sequencing. Clustalx-2.1-win, PolyPhen-2 and Swiss-PDBViewer software were used to analyze the conservatism of missense variation sites, whether the variations were harmful and their influences on protein structure and function. MutationTaster and NetGene2 software were used to analyze whether the splice site variation was harmful and its effect on the splice site. Results: The PT and APTT of the proband prolonged to 24.0 s and 69.8 s, respectively. The FⅤ: C and FⅤ: Ag decreased to 6% and 9%, respectively. There were compound heterozygous variations in F5 gene, which included c.911G>A heterozygous missense variation in exon 6 leading to p.Gly276Glu variation and c.5208+1G>A heterozygous missense variation in intron 15. The father and daughter had the p.Gly276Glu heterozygous variation. Her mother and son had the c.5208+1G>A heterozygous variation. Software analysis results of p.Gly276Glu heterozygous variation showed that Gly276 was conserved among homologous species, the variation was harmful, and it could affect the local structure and function of the protein. The c.5208+1G>A heterozygous variation was deleterious and resulted in the disappearance of the splice site, thereby affecting the protein function. Conclusion: The p.Gly276Glu and c.5208+1G>A compound heterozygous variants are deleterious variants associated with the patient's disease and may be the molecular pathogenesis of inherited FⅤ deficiency in this family.

Bioconversion of Hg0 into HA-Hg for simultaneous removal of Hg0 and NO in a denitrifying membrane biofilm reactor.

Bacterial mercury oxidation coupled to denitrification offers great potential for simultaneous removal of elemental mercury (Hg0) and nitric oxide (NO) in a denitrifying membrane biofilm reactor (MBfR). Four potentially contributory mechanisms tested separately, namely, membrane gas separation, medium absorption, biosorption and biotransformation, which contributed 4.9%/7.2%, 8.1%/8.9%, 38.8%/9.5% and 48.2%/84.9% of overall Hg0/NO removal in MBfR. Herein, Hg0 bio-oxidation, oxidative Hg0 biosorption and denitrification played leading roles in simultaneous removal of Hg0 and NO. Living microbes performed simultaneous Hg0 bio-oxidation and denitrification, in which Hg0 as electron donor was biologically oxidized to oxidized mercury (Hg2+), while NO as the terminal electron acceptor was denitrified to N2. The Hg2+ further complexed with humic acids in extracellular polymeric substances via functional groups (-SH, -OH, -NH- and -COO-) and formed humic acids bound mercury (HA-Hg). Non-living microbial matrix performed oxidative Hg0 biosorption, in which Hg0 may be physically adsorbed by cellular matrix, then non-metabolically oxidized to Hg2+ via oxidative complexation with -SH in humic acids and finally cleavage of S-H bond and surface charge transfer led to formation of HA-Hg. Therefore, bioconversion of Hg0 to HA-Hg by Hg0 bio-oxidation and oxidative Hg0 biosorption coupled with NO denitrification to N2 dynamically cooperated to accomplish simultaneous removal of Hg0 and NO in MBfR.

MiR-203 regulates DJ-1 expression and affects proliferation, apoptosis and DDP resistance of pancreatic cancer cells.

OBJECTIVE: DJ-1 is a negative regulator of PTEN and plays a role in tumorigenesis. Abnormal miR-203 expression is associated with pancreatic cancer. Bioinformatics analysis showed a targeted relationship between miR-203 and DJ-1 3'-UTR. This study investigated whether miR-203 regulates DJ-1 expression and its role in pancreatic cancer cell proliferation, apoptosis, and cisplatin (DDP) resistance. MATERIALS AND METHODS: The Dual-Luciferase reporter gene assay validated the targeted regulation between miR-203 and DJ-1. The DDP-resistant cell line SW1990/DDP was established and divided into miR-NC group and miR-203 mimic group followed by analysis of the expression of DJ-1, PTEN and p-AKT, cell apoptosis, and proliferation. RESULTS: There was a targeted relationship between miR-203 and DJ-1 mRNA. The expression of miR-203 in SW1990/DDP cells was significantly lower than that in SW1990 cells, while the expression of DJ-1 mRNA and protein was significantly higher than that in SW1990 cells. Compared with miR-NC group, the expression of DJ-1 and p-AKT protein in SW1990/DDP cells was significantly decreased in miR-203 mimic transfection group, while the expression of PTEN was significantly increased with increased cell apoptosis and decreased cell proliferation, as well as reduced DDP resistance. CONCLUSIONS: The decreased expression of miR-203 and the increased expression of DJ-1 is associated with drug resistance in pancreatic cancer cells. Elevated miR-203 can inhibit the expression of DJ-1, affect the activity of PTEN-PI3K/AKT pathway, inhibit the proliferation of pancreatic cancer cells, induce cell apoptosis, and reduce DDP resistance of pancreatic cancer cells.

Photocatalytic membrane combined with biodegradation for toluene oxidation.

Photocatalytic membrane coupled to biodegradation offers potential for degrading volatile organic compounds (VOCs) in photocatalytic membrane biofilm reactor. An intimately coupled photocatalysis and biodegradation reactor was operated in continuous operation for 500 days to treat simulated waste gas containing toluene. Toluene removal efficiency obtained 99%, with the elimination capacity of 550 g m-3·h-1. Membrane photocatalysis coupled to biodegradation was created to improve toluene removal from 11 to 20%. The dominant genera were Lysinibacillus, Hydrogenophaga, Pseudomonas at 30 d, Rudaea, Dongia, Litorilinea at 230 d xyl, Tod, Tcb, Bed, Tmo, Tbu, Tou, Dmp, Cat were functional genes of toluene metabolism, as shown by16S rDNA and metagenomic sequencing. Photocatalysis destroyed part of the toluene into biodegradable intermediates that were immediately mineralized by microorganisms in biofilm, some toluene was directly degraded by toluene degrading bacterial community into carbon dioxide and water. The novel hybrid photocatalytic membrane biofilm reactor is a cost-effective and robust alternative to VOCs treatment.

[Analysis of the influential factors of late diagnosis among newly identified HIV/AIDS cases in Anhui Province, 2011-2015].

Objective: To analyze the base situation and influential factors of late diagnosis among newly identified HIV/AIDS cases in Anhui Province from 2011 to 2015. Methods: Database information of the newly identified HIV/AIDS cases in Anhui Province from 2011 to 2015 were downloaded from the National HIV/AIDS Comprehensive Information System of China's disease prevention and control information system. To analyze the data including basic information, sample source, route of HIV transmission, population mobility, venereal disease, death and first CD4 count; and the number of 7 073 cases were classified according to late diagnosis and non-late diagnosis criteria. The Chi-square test and logistic regression analysis were used to analyze the influential factors of HIV late diagnosis. Results: A total of 7 073 newly identified HIV/AIDS cases were analyzed, and the mean age was (38.5±15.0) years. The proportion of late diagnosis in all counted cases was 41.7% (2 949/7 073); from 2011 to 2015, the proportions of late diagnosis were 59.7% (485/812), 46.5% (531/1 141), 42.7% (587/1 376), 36.1% (609/1 686), and 35.8% (737/2 058), respectively. Compared with the 0 to 19 years group, the 40 to 59 years group and over 60 years old group have higher risk of late diagnosis (OR=2.68, 95%CI: 1.94-3.71; OR=2.18, 95%CI: 1.53-3.10, respectively). Compared with the high education group, the illiterate and primary school education group have higher risk of late diagnosis (OR=1.74, 95%CI: 1.36-2.22; OR=1.64, 95%CI: 1.34-2.01, respectively). Compared with other sample sources, medical institutions have higher risk of late diagnosis (OR=2.64, 95%CI: 2.28-3.05). Compared with migrant population, the resident population have higher risk of late diagnosis (OR=1.80, 95%CI: 1.53-2.11). Conclusion: The proportion of late diagnosis among newly identified HIV/AIDS cases in Anhui province was relatively high from 2011 to 2015. The main risk factors of late diagnosis included cases reported by medical institutions, resident population, over 40 years old age group and low education level.

[A survey of correlation between serum 25-hydroxyvitamin D levels and dyslipidemia rlsk among middle-aged individuals in Beijing].

OBJECTIVE: To evaluate vitamin D status in middle-aged subjects in Beijing and explore the correlation between serum 25-hydroxyvitamin D[25(OH)D] levels and dyslipidemia. METHODS: A total of 448 individuals over 40 years old were enrolled in the cross-sectional survey. The general information, blood biochemical and lipid profiles and serum 25(OH)D levels were collected. The subjects were either divided into two groups (the dyslipidemia group and the non-dyslipidemia group) based on the lipid levels, or four groups according to quartiles of 25(OH)D levels. The association between 25(OH)D levels and dyslipidemia risk was analyzed by a logistic regression analysis. RESULTS: A total of 234 cases were in dyslipidemia group, which accounted for 52.23% of the subjects. The serum 25(OH)D levels were significantly lower in the dyslipidemia group than in the non-dyslipidemia group both in men and in women (all P<0.05). The median serum 25(OH)D level in the total subjects was 15.7 (12.2, 20.1)µg/L with 91.1% subjects of serum 25(OH)D level<30 µg/L. The proportion of subjects with dyslipidemia (high TC, high TG, high LDL-C, or low HDL-C) increased with the decrease of 25(OH)D level quartiles (P<0.05). After adjustment of confounding factors, the logistic regression analysis showed that subjects in the lowest 25(OH) D quartile group had 143% higher risks for dyslipidemia than those in the highest quartile group. CONCLUSION: These findings indicate that 25(OH)D insufficiency is highly prevalent among middle-aged individuals and it may be associated with the risk of dyslipidemia.

Expression of microRNA miR-34a inhibits leukemia stem cells and its metastasis.

OBJECTIVE: To study the expression profile and role of microRNA miR-34a in Leukemia stem cells (LSC) and during its metastasis. We examined upon the enforced expression of miR-34a in TIM3 positive leukemia stem cells and study the impact of leukemia stem cells and its metastasis. PATIENTS AND METHODS: Samples were collected from acute myeloid leukemia (AML) children along with controls. Immunohistochemistry analysis was performed with TIM3 antibody followed by Sorting of TIM3 positive cells by flow cytometry and the expression level of miR-34a was quantified by qRT-PCR assay. RESULTS: Immunohistochemistry in accordance with Sorting of TIM3 positive cells by flow cytometry concludes that the leukemia stem cells (LSC) were present in the samples. Induced expression of miR-34a in TIM3 positive leukemia stem cells (LSC), inhibits the clonogenic expansion, tumor progression and metastasis of leukemia. CONCLUSIONS: The enforced expression of miR-34a in TIM3 positive leukemia stem cells inhibits the leukemia stem cells and its metastasis. Our study illustrates that miR-34a is a key regulator and which will be developed as a novel therapeutic agent against leukemia stem cells (LSC).

[Correlation of serum sex hormone levels with metabolic syndrome in elderly men].

OBJECTIVE: To investigate the relationship between sex hormones and metabolic syndrome (MS), as well as its components in elderly men. METHODS: 1 505 elderly men (≥60 years old, mean age 75.4±9.7 years old) who participated in a routine health screening examination in PLA general hospital from May to June in 2012 were enrolled in this cross-sectional study. Serum lipids, glucose and sex hormones were measured along with body height, weight and blood pressure. Free testosterone (FT) and bioavailable testosterone (BT) were calculated. The correlation of serum sex hormones with the presence of MS and its components were analyzed. RESULTS: The prevalence of MS was 21.7% (326/1 505) in this study. Elderly men with MS had lower levels of sex hormone-binding globulin (SHBG), total testosterone (TT), FT and BT than those without MS. The levels of SHBG, TT, FT and BT were significantly lower in the overweight/obesity group, hyperglycemia group and dyslipidemia group than those in the respective control groups (P<0.05). Logistic regression analysis showed that the SHBG level was an independent risk factor for MS in elderly men(OR=0.977, 95%CI: 0.964-0.989, P<0.001), while the levels of TT, FT and BT were not associated with MS. The prevalence of MS gradually increased with decreasing of SHBG values (P<0.001). When comparing subjects in the lowest and highest quartile of SHBG, the former group demonstrated a 2.13-fold increase in the odds ratio for MS after adjusting for age, smoking, drinking and other sex hormone indices. CONCLUSION: In elderly men, lower SHBG level, not TT, FT or BT may be an independent predictor for the prevalence of MS, in which the mechanism requires further studies.

[Femtosecond lenticule extraction for correction of myopia: clinical results and recovery of subbasal nerves].

OBJECTIVE: To investigate the clinical efficacy, safety, predictability, corneal sensitivity, tear function and recovery of subbasal nerves after femtosecond lenticule extraction (FLEx) and laser in situ keratomileusis (LASIK). METHODS: In this prospective, nonrandomized, comparative clinical study, 49 patients (98 eyes) were divided into two groups. FLEx was performed to treat myopia by Visumax femtosecond laser system, and LASIK was performed by Allegretto Wave laser system. The patients were followed up for 6 months. Visual acuity, manifest refraction, intraocular pressure, slit-lamp examination, corneal topography by Pentacam, tear break-up time, Schirmer test, corneal sensitivity and confocal microscopy were assessed. RESULTS: Forty-four patients (88 eyes) completed the 6-month follow-up. Best corrected visual acuity (BCVA) was 0.89±0.14 in eyes with FLEx and 0.98±0.08 in eyes with LASIK at 1 day after surgery. After 10 days, BCVA was 0.98±0.09 and 1.02±0.09, respectively. At the final follow-up visit, the efficacy index was 1.09 in the FLEx group and 1.07 in the LASIK group, and the safety index was 1.12 and 1.07, respectively, in the two groups. Mean Schirmer score was (16.92±7.58) mm and (15.03±5.89) mm (t=1.316, P=0.192), mean tear break-up time was (8.94±2.57) s and (8.00±2.39) s (t=1.759, P=0.082), and corneal sensitivity was (56.46±4.49) mm and (51.38±8.16) mm (t=1.316, P=0.001) in the groups of FLEx and LASIK, respectively. At 10 days after surgery, the number of subbasal nerves was significantly decreased in the FLEx group, and in the LASIK group the subbasal nerve fibers were hardly observed. At 6 months, regenerated nerve fibers were evident in the subbasal area, which recovered faster in eyes with FLEx than in those with LASIK. CONCLUSIONS: Femtosecond lenticule extraction appears to be efficient, safe and predictable for myopia. FLEx surgery is superior over LASIK in less reduction of corneal sensation and lower risk of harm to the subbasal nerve fibers.

Effect of topiramate on apoptosis-related protein expression of hippocampus in model rats with Alzheimers disease.

OBJECTIVES: Alzheimer's disease is an age-related neurodegenerative disease and a synaptic function defect disease, the clinical symptoms are mainly progressive cognitive impairment, affecting patient's social function. Aim of this report was to investigate the effect of topiramate on apoptosis-related protein expression (Bcl-2, Survivin, Fas, Bax and Caspase-3) in the hippocampus of a rat model with Alzheimers Disease (AD). MATERIALS AND METHODS: Thirty-six adult Wistar rats were randomly divided into a control group, a model group and a test group. A dose of amyloid beta-protein 1-40 (Aß1-40) was injected into the hippocampus of the rats in the model and test groups, and the control rats are injected with same amount of saline. After AD model was successfully established, the rats in each group were administrated with an i.p. injection of topiramate (20 mg/kg/d) for 30 days. The effect of topiramate on the apoptosis-related protein levels in hippocampus neurons was studied by immunohistochemistry. RESULTS: The number of Bcl-2 and Survivin positive cells and optical density in hippocampus of rats in test group was more than those of rats in model groups, but less than those of rats in control group (p < 0.01); Fas, Bax and Caspase-3 positive cells and optical density in hippocampus of rats in test group was less than those of rats in the model group, but more than those of rats in control group (p < 0.01). CONCLUSIONS: Alzheimer's disease can induce apoptosis of hippocampus neurons in rats. Topiramate can prevent apoptosis of hippocampus neurons, at least in part, by increasing expression of Bcl-2 and Survivin and decreasing expression of Fas, Bax and Caspase-3.

Effect of edaravone on apoptosis of hippocampus neuron in seizures rats kindled by pentylenetetrazole.

OBJECTIVES: To explore the effect of edaravone (ED) on apoptosis of hippocampus neurons in seizures rats induced by pentylenetetrazole (PTZ). MATERIALS AND METHODS: Forty-eight adult Wistar rats were randomly divided into normal control (NC) group, PTZ group, and ED group. A dose of PTZ [35 mg/(kg·day)] was intraperitoneally (i.p.) injected into the rats of PTZ group and ED group until the kindling criterion was reached. After kindling, the rats of ED group were administered i.p. with ED [0.8 mg/(kg·day)]; the rats of PTZ group were administered i.p. with normal saline. After 30 min, seizures were induced by administering PTZ i.p. The influence of ED on Fas, Caspase-3, and Survivin protein immunoreactivity on hippocampus neurons was studied by immunohistochemistry method. RESULTS: Fas and Caspase-3 positive cells and optical density in hippocampus in PTZ group were more than that of ED group and NC group (all p < 0.01), but Survivin-positive cells and optical density in hippocampus in PTZ group were less than that of ED group (p < 0.01) and were more than that of NC group (p < 0.05); Fas- and Caspase-3-positive cells and optical density in hippocampus in ED group were more than that of NC group (all p < 0.05), but Survivin-positive cells and optical density in hippocampus in ED group were more than that of NC group (p < 0.01). CONCLUSIONS: Seizure can induce apoptosis of hippocampus neurons on seizures rats, but ED can resist the apoptosis of hippocampus neurons by increased expression of Survivin and decreased expression of Fas and Caspase-3.

Receptor-based QSAR study for a series of 3,3-disubstituted-5-aryl oxindoles and 6-aryl benzimidazol-2-ones derivatives as progesterone receptor inhibitors.

Receptor-based comparative molecular field analysis (CoMFA) and comparative molecular similarity indices analysis (CoMSIA) were performed on a series of 54 progesterone receptor (PR) inhibitors. The established CoMFA model from the training set gives statistically significant results with the cross-validated q (2) of 0.534 and non-cross-validated [Formula: see text] of 0.947. The best CoMSIA model was derived by the combination of steric field and hydrophobic field with a q (2) of 0.615 and [Formula: see text] of 0.954. A test set of 18 compounds was used to validate the predictive ability of the two models. The predicted correlation coefficients [Formula: see text] are 0.681 and 0.677 for CoMFA and CoMSIA models, respectively. Based on the CoMFA maps, the key structural characters of progesterone receptor inhibitors are identified. Moreover, the binding modes of oxindoles and benzimidazol-2-ones are also given by the quantum mechanical/molecular mechanical (QM/MM) calculations. This may provide useful information for drug design.

Upregulation of Fas and FasL expression in nicotine-induced apoptosis of endothelial cells.

Although smoking-related coronary vascular disease is well documented, the effects of nicotine have not been fully investigated. There is controversy over reports about the effect of nicotine on apoptosis. The effect of nicotine on apoptosis of human umbilical vein endothelial cells (HUVECs) and the expressions of Fas/Fas ligand (FasL) and caspase-3 were evaluated in this study. Annexin V fluorescein isothiocyanate and propidium iodide double staining demonstrated that nicotine (0.2 microM, 0.5 microM and 1 microM) could induce apoptosis of HUVECs; reverse transcription (RT)-PCR and Western blotting analysis demonstrated that levels of Fas and FasL expression were increased in nicotine-treated HUVECs. Moreover, caspase-3 expression was also increased. These data indicate that nicotine induces the apoptosis of HUVECs, and that the Fas/FasL pathway may play an important role. This provides evidence that nicotine may have an important role in cardiovascular pathology and atherogenesis.

Phylogenetic analysis of 16S rRNA gene sequences reveals rumen bacterial diversity in Yaks (Bos grunniens).

Six matured male Yaks (Bos grunniens) with a mean live weight of 450 +/- 23 kg (mean +/- SD), were housed indoors in metabolism cages and fed pelleted lucerne (Medicago sativum). After an adjustment period of 24 days of feeding the diet, samples of rumen content were obtained for analysis of the bacteria in the liquor. The diversity of rumen bacteria was investigated by constructing a 16S rRNA gene clone library using the general bacterial primers F27 and R1492. A total of 130 clones, comprising nearly full length sequences (approx. 1.5 kb) were sequenced and submitted to BLAST and phylogenetic analysis. Using the criterion that similarity of 97% or greater with the sequences of cultivated bacteria, 16 clones were identified as Butyrivibrio fibrisolvens, Pseudobutyrivibrio ruminis, Ruminococcus flavefaciens, Succiniclasticum ruminis, Selenomonas ruminantium and Prevotella ruminicola, respectively. A further 10 clones shared similarity ranging from 90 to 97% with cultivated bacteria but the similarity in sequences for the remaining 104 clones were less than 90% of those of cultivated bacteria. Using a phylogenetic analysis it was found that the majority of the clones identified (63.8%) were located in the Low G + C Subdivision, with most of the remainder (35.4% of clones) located in the Cytophaga-Flexibacter-Bacteroides phylum and one clone (0.8%) was identified as a Proteobacteria. It was apparent that Yaks have a large and diverse range of bacteria in the rumen content which differ from those of cattle and other ruminants.

Acetylcholinesterase: mechanisms of covalent inhibition of H447I mutant determined by computational analyses.

The reaction mechanisms of two inhibitor TFK(+) and TFK(0) binding to H447I mutant mouse acetylcholinesterase (mAChE) have been investigated by using a combined ab initio quantum mechanical/molecular mechanical (QM/MM) approach and classical molecular dynamics (MD) simulations. TFK(+) binding to the H447I mutant may proceed with a different reaction mechanism from the wild-type. A water molecule takes over the role of His447 and participates in the bond breaking and forming as a "charge relayer". Unlike in the wild-type mAChE case, Glu334, a conserved residue from the catalytic triad, acts as a catalytic base in the reaction. The calculated energy barrier for this reaction is about 8kcal/mol. These predictions await experimental verification. In the case of the neutral ligand TFK(0), however, multiple MD simulations on the TFK(0)/H447I complex reveal that none of the water molecules can be retained in the active site as a "catalytic" water. Taken together our computational studies confirm that TFK(0) is almost inactive in the H447I mutant, and also provide detailed mechanistic insights into the experimental observations.

[Analysis of organic additives in copper-plating brightener by high performance liquid chromatography].

This paper describes a high performance liquid chromatographic method for analysis of organic additives in copper-plating brightener. The copper-plating brightener was filtered through 0.45 microgram micropore membrane and determined on a Spherisorb-C6H5 column, with V(methanol):V(water) = 30:70 mobile phase at the flow rate of 1.5 mL/min, and detected at UV 210 nm. 2-benzimidazolethiol, ethylene thiourea, 1-propanesulfonic acid 3,3'-dithio-bis-disodium salt were separated and determined. The recoveries were more than 99% with CV less than 1%. The method is simple and rapid.