Purpose Focusing on the characteristics of for-malin fixed paraffin embedded(FFPE)samples,explored nano-magnetic bead nucleic acid extraction solutions with higher qual-ity/yield and continued to improve molecular pathology technolo-gy.Methods Alternative magnetic beads were synthesised in four major categories and 15 sub-categories and we screened to obtain high-quality/yield magnetic beads centred on FFPE samples.Simulated conventional tissues,simulated coarse needle punctures(liver),and simulated fiberoptic bronchoscopy sam-ples(lungs)were sectioned with the same number of serial slices in tubes.The nucleic acids of slices were extracted using the best magnetic beads screened in this study and common com-mercially available kits,and then perform comparison and purifi-cation quality parameters such as total amount and fragment size.The downstream applications of nucleic acids were validated by PCR and Sanger sequencing.Results Screening all homemade nanomagnetic beads centered on the DNA of FFPE samples,the total recoveries of the best performance nanomagnetic beads were obtained to be 58.5％±1.58％,and the total recoveries of five commercially available commercial magnetic beads and three do-mestic kit magnetic beads ranged from 18.68％to 40.71％.The total amount of DNA(ng)extracted from the same amount of tis-sue(serial slices),the nucleic acid yield of this study in simu-lated conventional tissues,simulated coarse needle punctures,and simulated fiberoptic bronchoscopy samples were increased by 39.49％-181.72％compared with those of the commercially a-vailable kits(P＜0.05).The total amount of extracted nucleic acid from simulated fiberoptic bronchoscopy tissue sections can be more than 100 ng for 1 slice(4 μm)and more than 400 ng for 5 slices.Conclusion The DNA purification nanomagnetic beads screened with DNA from FFPE samples have a significant enhancement comparing to the existing commercial bead proto-cols,and provide space for quality assurance,automated testing,and program expansion for clinical molecular pathology testing.

OBJECTIVES: This work aimed to investigate the molecular mechanism of cyclic tensile stress (CTS) stimulating autophagy in human periodontal ligament cells (hPDLCs). METHODS: hPDLCs were isolated and cultured from normal periodontal tissues. hPDLCs were loaded with tensile stress by force four-point bending extender to simulate the autophagy of hPDLCs induced by orthodontic force du-ring orthodontic tooth movement. XMU-MP-1 was used to inhibit the Hippo signaling pathway to explore the role of the Hippo-YAP signaling pathway in activating hPDLC autophagy by tensile stress. The expression levels of autophagy-related genes (Beclin-1, LC3, and p62) in hPDLCs were detected by real-time quantitative polymerase chain reaction. Western blot was used to detect the expression levels of autophagy-related proteins (Beclin-1, LC3-Ⅱ/LC3-Ⅰ, and p62) and Hippo-YAP pathway proteins (active-YAP and p-YAP) in hPDLCs. Immunofluorescence was used to locate autophagy-related proteins (LC3-Ⅱand p62) and Hippo-YAP pathway proteins (active-YAP) of hPDLCs. RESULTS: CTS-activated autophagy in hPDLCs and expression of autophagy-related proteins initially increased and then decreased; it began to increase at 30 min, peaked at 3 h, and decreased (P<0.05). CTS increased the expression of active-YAP protein and decreased the expression of p-YAP protein (P<0.05). When XMU-MP-1 inhibited the Hippo-YAP signaling pathway (P<0.05), active-YAP protein was promoted to enter the nucleus and autophagy expression was enhanced (P<0.05). CONCLUSIONS The Hippo-YAP signaling pathway is involved in the regulation of autophagy activation in hPDLCs under CTS.
Humans ; Hippo Signaling Pathway ; Periodontal Ligament/metabolism* ; Beclin-1/metabolism* ; Cells, Cultured ; Autophagy

OBJECTIVES: Since the outbreak of coronavirus disease 2019 (COVID-19), it has spread rapidly in China and many other countries. The rapid increase in the number of cases has caused widespread panic among people and has become the main public health problem in the world. Severe patients often have difficult breathing and/or hypoxemia after 1 week of onset. A few critically ill patients may not only rapidly develop into acute respiratory distress syndrome, but also may cause coagulopathy, as well as multiple organs failure (such as heart, liver and kidney) or even death. This article is to analyze the predictive role of clinical features in patients with COVID-19 for severe disease, so as to help doctor monitor the severity-related features, restrain the disease progress, and provide a reference for improvement of medical treatment. METHODS: The clinical data of 208 patients with COVID-19 who were isolated and treated in Changsha Public Health Treatment Center from January 17, 2020 to March 14, 2020 were collected. All patients were the mild and ordinary adult patients on admission, including 105 males and 103 females from 19 to 84 (median age 44) years old. According to the "Program for the diagnosis and treatment of novel coronavirus (COVID-19) infected pneumonia (Trial version 7)" issued by the General Office of National Health Committee and Office of State Administration of Traditional Chinese Medicine as the diagnostic and typing criteria. According to progression from mild to severe disease during hospitalization, the patients were divided into a mild group (=183) and a severe transformation group (=25). The clinical features such as age, underlying disease, blood routine, coagulation function, blood biochemistry, oxygenation index, and so on were analyzed. Among them, laboratory tests included white blood cell (WBC), lymphocytes (LYM), neutrophil (NEU), hemoglobin (Hb), platelet (PLT), prothrombin time (PT), plasma fibrinogen (Fib), activated partial prothrombin time (APTT), thrombin time (TT), -dimer, total bilirubin (TBIL), albumin (ALB), alanine aminotransferase (ALT), aspartate aminotransferase (AST), blood urea nitrogen (BUN), serum creatinine (Cr), creatine kinase (CK), creatine kinase isoenzyme-MB (CK-MB), lactate dehydrogenase (LDH), C-reactive protein (CRP), and oxygen partial pressure in arterial blood. Partial pressure of oxygen in arterial blood/fractional concentration of inspiratory oxygen (PaO/FiO) was calculated. The variables with statistical significance were analyzed by logistic regression analysis. RESULTS: Patients in the severe transformation group had more combined underlying diseases than those in the mild group (<0.05). From the perspective of disease distribution, patients in the severe transformation group had more combined hypertension (<0.05). In the severe transformation group, PT was significantly longer, the levels of Fib, ALT, AST, CK, LDH, and CRP were significantly higher than those in the mild group (<0.05 or <0.001), while LYM, ALB, and PaO/FiO were significantly lower than those in the mild group (<0.05 or <0.001). Logistic regression analysis was performed on clinical features with statistically significant differences. Combined with hypertension, LYM, PT, Fib, ALB, ALT, AST, CK, LDH, and CRP as independent variables, and having severe disease or not was the dependent variable. The results show that combined hypertension, decreased LYM, longer PT, and increased CK level were independent risk factors that affected the severity of COVID-19 (<0.05). CONCLUSIONS The patients with mild COVID-19 who are apt to develop severe diseases may be related to combined hypertension, decreased LYM, and longer PT, and increased CK level. For the mild patients with these clinical features, early intervention may effectively prevent the progression to severe diseases.
Adult ; Aged ; Aged, 80 and over ; Betacoronavirus ; China ; Coronavirus Infections ; diagnosis ; physiopathology ; Disease Progression ; Female ; Hospitalization ; Humans ; Male ; Middle Aged ; Pandemics ; Pneumonia, Viral ; diagnosis ; physiopathology ; Retrospective Studies ; Young Adult

Objective:To survey the status quo of ADR occurred in our hospital. Methods:Totally 277 ADR reports were collect-ed and analyzed statistically in our hospital from January 2010 to June 2013. Results:Among the ADR cases,the ratio of male to fe-male was 1∶1. 565, and 59. 93% of ADR occurred in the people over 50 years old. Totally 240 ADR cases (86. 64%) were induced by intravenous administration and 32 ADR cases (11. 87%) were induced by oral administration. Totally 167 ADR cases were caused by antibiotics (60. 29%), among them, 54 cases wero by cephalosporins (32. 34%), 47 cases by quinolones (28. 14%) and 32 ca-ses by cephamycins (19. 16%). Totally 42 ADR cases were caused by traditional Chinese medicine injections (15. 16%), and 18 ca-ses were caused by cardiovascular system drugs (6. 5%). The main clinical manifestations were lesion of skin and its appendant organr and digestive system injury with 162 cases (58. 48%) and 62 cases (22. 38%), respectively. Conclusion: It is necessary to strengthen clinical ADR monitoring, rationally use antibiotics and emphasize the importance of safe medication of traditional Chinese medicine injections.

Objective To determine the extracellular enzymatic activity of common pathogens for onychomycosis, in the hope of finding virulence factors associated with the pathogenesis of onychomycosis. Methods Strains tested in this study included standard strains of common dermatophyte and non-dermatophyte fungi as well as clinical isolates of Trichophyton rubrum from patients with onychomycosis. All the tested strains were cultured in medium containing nail fragments at 25 ℃ for 10 to 21 days followed by the determination of the nail fragment-containing medium, a significant increase was observed in the activities of esterase, esterase lipase, N-acetyl-β-glucosaminidase and α-mannosidase in dermatophytes compared with non-dermatophytes (all P ＜ 0.05 ), as well as in the activity of N-acetyl-β-glucosaminidase in Trichophyton rubrum compared with the other tested species of fungi (all P ＜ 0.05). No significant difference was noted in the activity of extracellular enzymes, except for that of naphthol-AS-BI-phosphohydrolase, between the isolates of Trichophyton rubrum from patients with different ranges of scoring clinical index for onychomycosis (SCIO). Conclusions In specific conditions, the extracellular enzymatic activity of fungi isolated from patients with onychomycosis is associated with fungal species, and may have a certain influence on the manifestations of anychomycosis.

A 19-year-old man was admitted to the hospital for erythema and nodules on the face and postauricular region for 6 years. Microscopic examination of lesion scrapings revealed brown septate hyphae. A restricted, velvety and black colony grew on Sabouraud's dextrose agar. Slide culture on potato dextrose agar plate showed flask-shaped phialides at the apex of or around the hyphae with clear collarettes and flaring apex,mucilage-encapsuled, round to oval, semi-endogenous phialosporae accumulating at the apex of the phialides,giving a flower-like appearance. Anti-fungal susceptibility test showed that the fungus was sensitive to itraconazole, terbinafine and amphotericin B, but resistant to fluconazole. Sequence analysis of the ITS1-ITS4 region revealed a 98% consistency with the reference sequence of ITS1-ITS4 of Phialophora verrucosa. On the basis of above findings, the patient was diagnosed with cutaneous phaeohyphomycosis. Clinical improvement was seen after treatment with oral itraconazole (400 mg/d).

Objective To explore the value of echocardiography in the diagnosis of the criss-cross heart. Methods The echocardiographic results of 6 patients with the criss-cross heart were reviewed retrospectively. The echocardiographic characteristics were analyzed and compared with the surgery results in 4 operated cases. Results The 6 cases were interpreted as representing a criss-cross heart with solitus atria,D-loop ventricles and concordant atrioventricular connections. The ventriculo-arterial alignments of 5 cases were abnormal including double outlet right ventricle with anterior aorta in 2 and transposition of the great arteries in 3 while 1 patient had concordant connection. The following principal characteristics were tilting the transducer from posterior to anterior could demonstrate the connection of the left-sided left atrium and the right-sided left ventricle through mitral valve at first. The more anterior angulation of the transducer then showed the right-sided right atrium was connected to the left-sided right ventricle through tricuspid imaging displayed the two atrial outflows crossed each other without mixing at atrioventricular valve level.right ventricle often occured. The echocardiographic diagnosis of the criss-cross heart and its associated cardiac abnormalities were confirmed by surgery in 4 cases except 1 persistent left superior vena cava was missed. Conclusions The invisibility of a standard 4-chamber view in any cut was very characteristic in the echocardiographic diagnosis of the criss-cross heart. The definitive appearance was the separate display of the two ventricular inlets and the crossed atrioventricular connections with each atrium emptying into the contralateral ventricle by continuous subxiphoid or apical scanning. The transthoracic echocardiography can diagnose this rare heart disease and associated cardiac abnormalities accurately.

A 16-year-old woman presented plaques on the left auricle and face over a period of 3 years. Fungal culture grew black-grey or dust velvety colony on Sabouraud's dextrose agar plate. A slide culture on potato dextrose agar plate showed conidiophores which were unbranched or occasionally loosely branched. The conidia were sympodial, zero- to two- septate, with rounded apices and truncated bases. The optimum growth temperature was 26℃ - 30℃. The fungus had the ability to liquefy glutin and hydrolyze starch. Anti-fungal susceptibility test showed the fungus was susceptible to itraconazole, terbinafine and amphoterecin B, but resistant to fluconazole. Cutaneous biopsy specimens revealed brown hyphae and budding yeast cells. The sequence of internal transcribed spacer (ITS) 1-ITS4 region of the isolate rDNA was assessed and compared against the Genebank databases. A 99% consistence was observed in the ITS sequence between clinical isolate and reference strain of Veronaea botryose Ciferri et Momtemartini. Based on the above findings, the mold was identified as Veronaea botryose Ciferri et Momtemartini. The lesions gradually subsided after 8-month treatment with oral itraconazole of 100 mg twice daily.

Objective To investigate the effect of electron transfer system on the hyphal formation of Candida albicans. Methods Candida albicans was cultured in RPMI 1640 supplemented with 10% new-born calf serum in 5% CO2 at 37 ℃ with or without the presence of inhibitors or activators of electron transfer system. Growth curve, morphology and percent of filamentation were observed for Candida albicans. MTT assay was used to assess the viability of Candida albicans. Results The solvents (chloroform and dimethyl sulfoxide) had no significant effect on the growth of and filamentation in Candida albicans. After incubation with thenoyltrifluoroacetone (TTFA) or benzhydroxamic acid for 24 hours, yeast cells of Candida albicans predominated in the culture. The growth of Candida albicans was significantly inhibited in log phase by the incubation with classic respiratory chain inhibitors such as rotenone, antimycin A, oligomycin, sodium azide, TTFA and sodium malonate, compared with the controls (all P < 0.01). Benzhydroxamic acid, an inhibitor of alternative oxidative pathway, also significantly inhibited the growth of Candida albicans in log phase (t = 10.92, P < 0.01). After incubation with rotenone, antimycin A, oligomycin, sodium azide, TTFA, sodium malonate, benzhydroxamic acid and disodium gnanylate, the percentage of filamentation in Candida albicans at 12 hours was 87.49 ± 0.52, 48.75 ± 4.44, 50.33 ± 8.50, 99.00 ± 1.00, 1.60 ± 0.53, 94.01 ± 0.99, 0.00 ± 0.00 and 92.33 ± 2.08, respectively, and the growth of Candida albicans at 7 hours was inhibited by (1.34 ± 0.15)%, (70.61 ± 1.02)%, (50.63 ± 5.38)%, (17.80 ± 7.89)%, (45.17 ± 1.27)%, (10.75 ± 3.62)%, (72.46 ± 1.14)% and -(5.96 ± 4.07)%, respectively. Conclusions Hyphal formation of Candida albicans could be suppressed by inhibitors of classic respiratory chain or alternative oxidative pathway, and is mainly regulated by alternative oxidative pathway.

Objective To analyze the left ventricular(LV) rotation and twist by two-dimensional ultrasound speckle-tracking imaging(STI),and evaluate the correlation between twist and other components of LV contraction in uremia patients. Methods Twenty-nine uremia patients and twenty-nine healthy volunteers were included. High frame rate two-dimensional images were recorded from the left ventricular short-axis views both of the basal and apical planes. Segmental and global LV rotation were recorded at both The peak systolic LV twist was significantly decreased in uremia patients(P <0. 001). The loss of LV twist occurred mainly by the impairment of maximal global counterclockwise apical rotation(P <0. 001),and was augmented by somewhat less reduction in global clockwise basal rotation although the differences was not significantly reduced(P<0.01), while only the posterior and inferior of the basal plane was significantly fraction(r=0.645,P=0.000), LV end-diastolic volume (r=-0.549,P=0.002), LV end-systolic volume(r=-0.673, P = 0.000) in uremia patients. Conclusions STI enables quantification of segmental and global LV rotation in an objective manner in uremia patients,and provides a new method for evaluation of segmental and global LV systolic function.