[Classification of cardiac amyloidosis: an immunohistochemical analysis].

Objective: To evaluate the sensitivity and specificity of immunohistochemistry (IHC) in the classification of cardiac amyloidosis on endomyocardial biopsy (EMB) and heart allograft. Methods: Twenty cardiac tissues from 19 patients at Fuwai Hospital from January, 1990 to April, 2017 with histopathologic features of amyloidosis and Congo red staining positivity were included. IHC was performed with monoclonal antibodies against AA amyloid and polyclonal antibodies against transthyretin (ATTR), λ-light chain (AL-λ), κ-light chain (AL-κ), ApoAⅠ, ApoAⅡ, ApoA Ⅳ and ß(2)-microglobin. The extent of interstitial staining was evaluated by light microscopy, and three patterns were recognized; these included diffuse pericellular pattern, discrete pericellular pattern, and nodular pattern. Two patterns of vascular deposition were also noted, including arterial pattern and venous pattern. Endocardial involvement was also assessed and recorded. Results: Nineteen cases were divided into three groups according to the pattern of proteins expression in specimens. The first group (5 cases) only showed single protein expression on EMB. The second group (6 cases) showed more than one protein expression, but one of them was intensely stained or any staining of any protein together with ApoA Ⅳ co-staining. The third group (8 cases) also showed more than one protein expression and all of them had intense staining. Amyloid deposits were successfully subtyped as AL-λ, ATTR, AL-κ and ApoAⅠby IHC in the former two groups with the sensitivity of 11/19. In the third group, amyloid deposits could not be subtyped by immunohistochemistry due to their poor specificity. The pericellular pattern tended to favor AL over ATTR amyloidosis and vascular deposition tended to favor ATTR. Conclusions: Amyloid deposits can be reliably subtyped in diagnostic cardiac specimens using IHC. The co-deposition of chaperon proteins, the distribution of amyloid proteins and clinical features are also auxiliary to subtype cardiac amyloidosis.

[The clinical characteristics, diagnosis and treatment of patients with gout in China].

Objective: To investigate the demographic characteristics, clinical features, diagnosis and treatment of patients with gout in China. Methods: Clinical data of 6 814 patients with gout from 100 hospitals in 27 provinces, municipalities or autonomous regions in China were collected and analyzed. Results: (1) The ratio of male to female in patients with gout was 14.7∶1. The mean age of onset was (48.8±15.1) years old. Mean serum urate level was (526.7±132.3) µmol/L. Patients' education background was of U-shaped distribution; (2) Hypertension was the most common comorbidity [15.8%(1 079/6 814)], then overweight or obesity [51.9%(3 536/6 814)]; (3) Alcohol and high-purine food intake were dominant triggering factors in men. The diagnosis of gout was made after onset in majority of patients with cardinal symptom arthralgia. Most patients had the disease less than 5 years, and the longer the course, the more flares in the previous year of entry; (4) Febuxostat was the mostly used urate-lowering medication. 20.7%(1 412/6 814), 10.8%(739/6 814) and 3.9%(265/6 814) of patients were followed up in 4 weeks, 12 weeks and 24 weeks after registration, and 18.9%(267/1 412), 29.1%(215/739) and 38.1%(101/265) of them reached the control target of serum urate levels, respectively. After treatment, patients' liver function was not affected, but serum creatinine levels decreased significantly. Conclusions: The proportion of gout patients who reach target serum urate level is very low. Further steps including education and survey need to be carried on.

[Analysis on clinical characteristics and prognosis of patients with mitral valve prolapse].

Objective: To evaluate the clinical characteristics and prognosis of patients with mitral valve prolapse (MVP). Methods: We retrospectively analyzed the clinical characteristics and prognosis of 148 MVP patients who underwent mitral valve surgery in Fuwai hospital from January 2012 to December 2015.The patients were divided into mucoid degeneration group (52 cases) and without mucoid degeneration group(19 cases) according to pathological examination of leaflets and chordate. Results: The clinical symptoms of MVP patients included dyspnea (59.5%(88/148)), chest distress and pain (52.7%(78/148)), and palpitations (36.5% (54/148)). Mitral valve repair was performed in 144 cases (97.3%), and mitral valve replacement was performed in 4 cases (2.7%). Posterior leaflet prolapse was the most common form of MVP (68.9%, 102/148). Pathological examination revealed myxomatous degeneration in 73.2% patients (52/71), fibrosis in 8.5% patients (6/71), and fibrinoid necrosis in 8.5% patients (6/71). Patients with mucoid degeneration had less atrial fibrillation before surgery (5.8%(3/52) vs. 42.1%(8/19), P<0.01), smaller preoperative left atrium diameter ((43.2±6.5) mm vs. (48.2±8.9) mm, P<0.05), more posterior leaflet prolapse (94.2%(49/52) vs. 63.2%(12/19), P<0.01), redundant chordae (26.9%(14/52) vs. 0, P<0.05) and leaflet thickening (76.9%(40/52) vs. 52.6%(10/19), P<0.05) when compared with patients without mucoid degeneration.Echocardiography examination at the postoperative follow-up of 39.0(22.3, 57.0) months revealed smaller left atrium diameter((38.5±7.1) mm vs. (45.3±8.3) mm, P<0.01), left ventricular end-diastolic diameter ((48.9±6.2) mm vs. (57.5±7.6) mm, P<0.01), reduced left ventricular ejection fraction ((61.2±7.1)% vs. (65.1±6.2)%, P<0.01) and less moderate or severe mitral regurgitation (1.4%(2/148) vs. 100.0%(148/148), P<0.01) compared with the corresponding preoperative values. Conclusions: Dyspnea is the main symptom, and mucoid degeneration characterized by redundant chordae and leaflet thickening are the main pathological features of MVP patients.The surgical treatment of MVP patients is related with satisfactory outcome results.

[Clinical significance of FOXM1 and Gli-1 protein expression in high-grade ovarian serous carcinoma].

Objective: To investigate the different expression and prognostic significance of forkhead box M1 (FOXM1) and Gli-l in ovarian high grade serous carcinoma (HGSC). Methods: The expressions of FOXM1 and Gli-1 in 94 cases of HGSC and 20 cases of normal fallopian tube tissues were detected by immunohistochemistry. Kaplan-Meier analysis and Cox multivariate survival analysis were used to assess the relationship of the FOXM1 and Gli-1 levels with age, International Federation of Gynecology and Obstetrics (FIGO) stage, omental metastasis, and residual foci and prognosis of HGSC. Results: The positive rates of FOXM1 and Gli-1 expression in HGSC were 79.8% (75/94) and 77.7% (73/94), respectively, both significantly higher than those of the normal controls (P<0.05). The expressions of FOXM1 and Gli-1 were significantly correlated with FIGO stage, and both of their positive rates in stage Ⅲ-Ⅳpatients were significantly higher than those in stage Ⅰ-Ⅱ cases (P<0.001). The expressions of FOXM1 in HGSC were positively correlated with Gli-1.Kaplan-Meier analysis revealed that the 5-year overall survival rates of FOXM1- and Gli-1-positive groups were 8.0% and 6.8%, significantly lower than 36.8% and 38.1% of the FOXM1- and Gli-1-negative groups, respectively (P<0.05 for both). Cox multivariate survival analysis revealed that FIGO stage and overexpression of FOXM1 protein were independent prognostic factors of HGSC patients (P<0.05 for both). Conclusions: The overexpression of FOXM1 and Gli-1 proteins participate in the carcinogenesis of HGSC, and are significantly associated with FIGO stage. The protein expression of FOXM1 is positively correlated with Gli-1 in HGSC. Expression of FOXM1 protein and FIGO stage are independent prognostic factors of HGSC.

[Surgical pathology analysis of the causes of failure of 48 bioprosthetic heart valves in 40 Chinese cases].

OBJECTIVE: To investigate the pathological changes and related factors leading to failure of bioprosthetic valves. METHODS: The surgical explanted bioprosthetic valves in Fuwai Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College from January 2006 to December 2014 were retrospectively reviewed. Calcification, cusp tears, pannus and infectious endocarditis of bioprosthetic valves were evaluated by gross examination, X ray photography and histopathological analysis. Meanwhile, the age and the site at implantation, and the tissue type of bioprosthetic valves, the implantation duration were obtained by chart review and used to be analyzed with the modes of valve failure. U test, χ(2) test and Fisher exact test were used to analyze the data. RESULTS: Total 48 bioprosthetic valves were collected from 40 cases with 26 females. Their age at the time of implantation was 12 to 71 years (median 58 years). The tissue types of bioprosthesis included 24 bovine pericardial valves and 24 porcine aortic valves respectively. The implantation sites included mitral in 29 valves, aortic in 14 valves and tricuspid in 5 valves. The implantation duration ranged from 5 days to 27 years (mean (111±77) months). The study showed that the commonest mode of valve failure was structural valve deterioration (32 valves, 66.7%). The fewer modes of valve failure were infectious endocarditis (9 valves, 18.7%) and non-structural valve deterioration (7 valves, 14.6%). No thrombosis was found in this series. The degree of calcification was correlated positively with implantation duration (r=0.52, P=0.000). The implantation duration of bovine pericardial valves was longer than that of porcine aortic valves ((146±74) months vs. (77±68) months, Z=-3.24, P=0.001). Porcine aortic valves had a higher incidence of cusp tears than bovine pericardial valves (66.7% vs. 16.7%, χ(2)=12.34, P=0.000) despite they had a relatively lower incidence of calcification (41.7% vs. 75.0%, χ(2)=5.37, P=0.019). For the age at the time of implantation, the patients ≤60 years old had more calcification of bioprosthetic valves compared with those >60 years old (45.1% vs. 2/17, χ(2)=5.51, P=0.019) whereas the patients ≤60 years old had fewer cusp tears (12.9% vs. 58.8%, P=0.002). Infectious endocarditis was found more frequently in the valves failure within 5 years than those failure after 5 years (6/14 vs. 8.8%, P=0.006). CONCLUSIONS: Structural valve deterioration is the principle cause of bioprosthetic valve failure while infectious endocarditis mainly involves the valves failure in early and middle terms after surgery. The mode of valve failure in the patients ≤60 years old is different from that in the patients >60 years old at the time of implantation. More calcification would occur in bioprosthetic valve in the younger patients.

A multiplex touchdown PCR for detection of Streptococcus pneumoniae, Haemophilus influenzae type b and Mycobacterium tuberculosis complex in sputum samples.

Rapid and accurate detection of Streptococcus pneumoniae (Sp), Haemophilus influenzae type b (Hib) and Mycobacterium tuberculosis complex (MTBC) in sputum by conventional methods remains problematic. Primers based on capsular polysaccharide biosynthesis gene (cpsA), the region II of the capsulation locus (cap), the insertion sequence IS6110 were designed for Sp, Hib, MTBC respectively. These primers were incorporated in a multiplex touchdown PCR assay for simultaneous detection of Sp, Hib and MTBC. The multiplex touchdown PCR assay was evaluated using standard strains and clinical sputum samples. The multiplex touchdown PCR assay showed 100% specificity in identifying Sp, Hib, MTBC from pure culture of standard strains. The sensitivities of the multiplex touchdown PCR assay were 94%, 98%, 98% for detection of Sp, Hib and MTBC respectively based on culture results while evaluated using 492 consecutive qualified clinical sputum samples; the specificities were all 100%. This highly sensitive and specific multiplex touchdown PCR assay offers a rapid and simple method for detection of Sp, Hib and MTBC in clinical sputum samples.

A multiplex touchdown PCR for detection of Streptococcus pneumoniae, Haemophilus influenzae type b and Mycobacterium tuberculosis complex in sputum samples

Rapid and accurate detection of Streptococcus pneumoniae (Sp), Haemophilus influenzae type b (Hib) and Mycobacterium tuberculosis complex (MTBC) in sputum by conventional methods remains problematic. Primers based on capsular polysaccharide biosynthesis gene (cpsA), the region II of the capsulation locus (cap), the insertion sequence IS6110 were designed for Sp, Hib, MTBC respectively. These primers were incorporated in a multiplex touchdown PCR assay for simultaneous detection of Sp, Hib and MTBC. The multiplex touchdown PCR assay was evaluated using standard strains and clinical sputum samples. The multiplex touchdown PCR assay showed 100% specificity in identifying Sp, Hib, MTBC from pure culture of standard strains. The sensitivities of the multiplex touchdown PCR assay were 94%, 98%, 98% for detection of Sp, Hib and MTBC respectively based on culture results while evaluated using 492 consecutive qualified clinical sputum samples; the specificities were all 100%. This highly sensitive and specific multiplex touchdown PCR assay offers a rapid and simple method for detection of Sp, Hib and MTBC in clinical sputum samples.

Enhanced osseointegration of titanium implant through the local delivery of transcription factor SATB2.

Titanium implants are widely used in dentistry and orthopedic surgery. Nevertheless, bone regeneration around the implant is a relatively slow process, after placement. This study assessed whether SATB2 can enhance osseointegration of a titanium implant. To determine the effect of SATB2 in implant integration, two different viruses encoding SATB2 (PBABE-Satb2 virus or RCAS-Satb2 virus) were locally administered to the bone defect prior to titanium implant placement in our established transgenic TVA mice. Seven and 21 days post implantation, the femurs were isolated for quantitative real-time RT-PCR, H&E staining, immunohistochemical (IHC) staining, and microcomputed tomography (microCT) analysis. Quantitative real-time RT-PCR results demonstrated that the in vivo overexpression of SATB2 enhanced expression levels of potent osteogenic transcription factors and bone matrix proteins. We also found that 21 days after implantation, there were no significant differences in the expression levels of SATB2, Osx, Runx2, COLI, OC, and BSP between the RCAS-Satb2 group and the RCAS group. Histological analysis showed that SATB2 overexpression significantly enhanced new bone formation and bone-to-implant contact after implantation. IHC staining analysis revealed that forced expression of SATB2 increased the number of BSP-positive cells surrounding the implant. MicroCT analysis demonstrated that in vivo overexpression of SATB2 significantly increased the density of the newly formed bone surrounding the implant. These results conclude that in vivo overexpression of SATB2 significantly accelerates osseointegration of titanium implants and SATB2 can serve as a potent molecule in promoting tissue regeneration.

Effect of cinnamaldehyde on biofilm formation and sarA expression by methicillin-resistant Staphylococcus aureus.

AIMS: To investigate the antibiofilm effect of cinnamaldehyde on methicillin-resistant Staphylococcus aureus (MRSA) and analyse the effect of subminimum inhibitory concentrations (MICs) of cinnamaldehyde on the expression of the biofilm-related gene sarA. METHODS AND RESULTS: The MICs and minimum bactericidal concentrations (MBCs) were determined using a microtitre broth dilution method. Biofilm susceptibility was determined using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) staining and colony forming unit (CFU) counting assays. Antibiofilm effects were studied with scanning electron microscopy (SEM) and confocal laser scanning microscopy (CLSM). SarA expression was assessed by real-time PCR. MICs and MBCs were in the range 0.0625-0.5% (v/v). The killing effects were concentration dependent. At a concentration of 5× MIC, all strains in biofilm were decreased to lower than 20% of the control groups. SEM and CLSM images indicated that a 5× MIC concentration of cinnamaldehyde was able to detach and kill existing biofilms. Apart from strain JB-06, real-time PCR showed that the expression of sarA of all other strains was decreased upon exposure to sub-MICs of cinnamaldehyde. CONCLUSIONS: These data showed the strong killing effect of cinnamaldehyde against MRSA within biofilms. SIGNIFICANCE AND IMPACT OF THE STUDY: This study indicated the potential of cinnamaldehyde as an inhibitory agent for use in MRSA biofilm-related infections.

Gene expression profile in the liver tissue of geese after overfeeding.

Geese form a fatty liver after feeding on a carbohydrate-rich diet, possibly as an evolutionary adaptation to accumulate reserves for migration. To gain insight into the gene-regulation processes of hepatic steatosis in geese, we examined the profile of transcriptional expression in goose fatty liver and control liver by suppression subtractive hybridization and measured the levels of serum biochemical variables. We found 107 genes whose expression was different between the treatment and control groups. The main functions of these genes are metabolic processes, including the metabolism of carbohydrates, amino acids, and lipids. Twenty-four genes were classified using the Kyoto Encyclopaedia of Genes and Genomes pathways. Twelve genes that related to metabolic and cellular processes were confirmed by quantitative RT-PCR. A specific positive effect of feeding was observed on the expression of genes involved mainly in unsaturated fatty acids and triglyceride synthesis, and a negative effect was observed on genes involved in ß-oxidation, cholesterol metabolism, and glycolysis. The results could serve as an important reference for the development of goose breeding for fatty liver production and human liver disease research.

Influence of magnetic field on Cr(VI) adsorption capability of given anaerobic sludge.

To provide beneficial guide for the application of the magnetic field in the bio-treatment of the Cr(VI)-contained wastewater, sludge samples from the control bio-system A (absent of magnetic field) and the contrast bio-system B (present of magnetic field) were used to adsorb the synthetic wastewater with 100 mg l(-1) Cr(VI). Influences of two adsorption modes, single adsorption and once continuous adsorption, on the Cr(VI) adsorption capacities of both sludge samples were compared. And the influence of regeneration on the Cr(VI) adsorption capacities were also studied. The results of adsorption experiments showed that the Cr(VI) adsorption capacities of the first single adsorption for sludge sample A and B were pretty nearly, which were 9.79 and 9.93 mg, respectively. And after 5 single adsorption periods, the total Cr(VI) adsorption capacity and efficiency of the sample B were 25.88 and 55.66 mg Cr(VI) g(-1)VSS, while those of the control were 14.95 and 33.98 mg Cr(VI) g(-1)VSS, respectively. For the sludge sample A and B after a single adsorption, both functions of regeneration were remarkable. But after 13 cycles of the single adsorption-regeneration, the Cr(VI) adsorption capacity and efficiency of the sample B were 110.15 and 189.91 mg Cr(VI) g(-1)VSS, while those of the control were 70.89 and 140.38 mg Cr(VI) g(-1)VSS, respectively. Though the Cr(VI) adsorption capacity of a once continuous adsorption period was more than that of a single adsorption period obviously, the Cr(VI) removal rates of the sludge sample A and B in the third period of once continuous adsorption-regeneration were only 8.12 and 33.51%, respectively. It was concluded that the weak magnetic field did improve the Cr(VI) bio-removal efficiency and the sludge stability, the batch treatment was an ideal operation mode for the bio-treatment of the Cr(VI)-contained wastewater, as compared with the continuous operation mode, but regeneration and enough sludge content were two necessary conditions to ensure the efficiency of batch treatment.

Influence of magnetic field on activity of given anaerobic sludge.

Two modes of magnetic fields were applied in the Cr(6+) removal sludge reactors containing two predominated strains--Bacillus sp. and Brevibacillus sp., respectively. The magnetic field mode I* of 0-4.5 or 0-14 mT between pieces was obtained by setting the magnetic pieces with the surface magnetic density of 0-6 or 0-20 mT into the reactor, and the magnetic field mode II* of 6, 20, or 40 mT on the return line was obtained by controlling the working distance of the permanent magnet outside the sludge return line. The effects of different magnetic fields on the activity of the given anaerobic sludge were studied by comparing with the control (absent of magnetic field). The results showed that the magnetic field of 0-4 mT improved the activity of given sludge most effectively, U(max) CH(4) (the peak methane-producing rate) and the methane producing volume per gCOD(Cr) reached 64.3 mlCH(4)/gVSS.d and 124 mlCH(4)/gCOD(Cr), which increased by 20.6 and 70.7%, respectively, compared with the control. And the magnetic field of 20 mT took second place. It could be concluded that the input of some magnetic field could improve the activity of anaerobic sludge by increasing the transformation efficiency of COD(Cr) matters to methane, and the total organic wastage did not increase.

A comparative study on SWCNT and DWCNT field-effect transistors.

Field-effect transistors have been fabricated using single-walled carbon nanotubes (SWCNTs) and double-walled carbon nanotubes (DWCNTs), and their electrical transport properties have been studied comparatively. While a semiconducting SWCNT exhibits better field-effect characteristics than a DWCNT counterpart, the DWCNT shows more complicated response to external gate modulation. Depending on the nature of the two shells of a DWCNT, i.e., whether the shell is semiconducting (S) or metallic (M), a DWCNT device can be described as either S-S, or S-M, or M-S, or M-M. It was found that the S-S and M-M or M-S devices show similar field-effect characteristics to those found in SWCNT devices. But for S-M DWCNT devices, distinct field-effect characteristic was found and attributed to the combined effects of intershell interactions and screening by free carriers of the inner metallic shell. The S-M DWCNT devices thus provide a perfect system for studying the important intershell interaction, and information on the effect of this interaction on the electrical properties of a multi-walled carbon nanotube can be obtained by a comparative study of S-M DWCNT and S-SWCNT devices.

DIP: The Database of Interacting Proteins: 2001 update.

The Database of Interacting Proteins (DIP; http://dip.doe-mbi.ucla. edu) is a database that documents experimentally determined protein-protein interactions. Since January 2000 the number of protein-protein interactions in DIP has nearly tripled to 3472 and the number of proteins to 2659. New interactive tools have been developed to aid in the visualization, navigation and study of networks of protein interactions.

Serotonin-like immunoreactive fibers in the medial dorsal accessory group and the anterior fornical nucleus of the magnocellular neurosecretory system in the rat.

The area lateral to the stria medullaris and the fornix at the level of the interventricular foramen, an area known to be occupied by the medial dorsal accessory group and the anterior fornical nucleus of the magnocellular neurosecretory system, was found to be studded with serotonin (5-HT)-like immunoreactive (LI) varicosities. Double immunostaining with antisera against 5-HT and oxytocin (OXY) proved that the 5-HT-LI varicosities were in close proximity to the OXY-LI neurons in the medial dorsal accessory group and the anterior fornical nucleus. These nuclei were distributed more densely with 5-HT-LI varicosities than other cell groups in the magnocellular neurosecretory system.