[Clinical phenotype and genetic analysis of patients with left ventricular noncompaction caused by the biallelic mutation of MYBPC3 and MYH7].

Objective: To explore the relationship between pathogenic gene, mutation and phenotype of left ventricular noncompaction (LVNC) patients and their family members. Methods: The subjects were the proband with LVNC and her family members. The medical history including electrocardiogram, echocardiography and cardiac magnetic resonance examination of the proband and family members were collected. Whole exome sequencing of the proband was performed, bioinformatics analysis focused on the genes related to hereditary cardiomyopathy. Candidate pathogenic sites were validated by Sanger sequencing. The clinical interpretation of sequence variants were classified according to American College of Medical Genetics and Genomics (ACMG) guidelines. Results: The proband carried a heterozygous variation of the MYBPC3 gene c.C2827T and the MYH7 gene c.G2221C. The proband's sister carried heterozygous variation of MYBPC3 gene c.C2827T. According to the ACMG guidelines, the variant was determined to be pathogenic. Conclusion: The missense variant of MYBPC3 gene c.C2827T and MYH7 gene c.G2221C are identified from the proband with LVNC and her family member, which provides a genetic basis for clinical diagnosis and genetic counseling of the patients and the family members with LVNC.

Optimal reference genes for real-time quantitative PCR and the expression of sigma factors in Acidithiobacillus caldus under various conditions.

AIMS: Acidithiobacillus caldus is an important sulphur-oxidizing bacterium that plays crucial roles in the bioleaching industry. This study aims to analyse the optimal reference gene for real-time quantitative PCR (RT-qPCR) under different conditions and investigate the transcription levels of the sigma factor genes in the stress response. METHODS AND RESULTS: We selected six housekeeping genes and analysed them via RT-qPCR using two energy resources, under four stress conditions. Three statistical approaches BestKeeper, geNorm, and NormFinder were utilized to determine transcription stability of these reference genes. The gapdH gene was the best internal control gene using elemental sulphur as an energy resource and under heat stress, map was the best internal control gene under pH and osmotic stress, era was the best internal control gene for the K2 S4 O6 energy resource, and rpoC was the best internal control gene under Cu2+ stress. Furthermore, the expressional levels of 11 sigma factors were analysed by RT-qPCR in the stress response. CONCLUSIONS: Stable internal control genes for RT-qPCR analysis of A. caldus were determined, and the expression patterns of sigma factor genes of A. caldus were investigated. SIGNIFICANCE AND IMPACT OF THE STUDY: The identification of the optimal reference gene and analysis of transcription levels of sigma factors in A. caldus can provide clues for reference gene selection and the study of sigma factor function.

Ultra-long high quality catalyst-free WO3 nanowires for fabricating high-performance visible photodetectors.

This work presents a study on the controlled growth of WO3 nanowires via chemical vapor deposition without catalyst, and their potential applications in visible photodetectors. The influence of growth conditions on the morphology of WO3 nanowires is studied in order to understand the growth mechanism of WO3 nanowires, and ultra-long (60 [Formula: see text], the longest one ever reported) WO3 nanowires with a spindle shape are achieved by optimizing the growth conditions. It was found that the length of WO3 nanowires increases from 15 [Formula: see text] to 60 [Formula: see text] with increasing the argon carrier gas flow rate from 30 sccm to 90 sccm, and then saturates with further increasing the argon carrier gas flow rate. However, the length of WO3 nanowires reduces from 60 [Formula: see text] to 19 [Formula: see text] with increasing the tube inner pressure from 2.5 Torr to 3.5 Torr. The photoconductor detectors based on WO3 single nanowires present excellent device performance with a responsivity as high as 19 A W-1 at a bias of 0.1 V, a detectivity as high as 1.06 × 1011 Jones, and a response (rising and decay) time as short as 8 ms under the illumination of a 404 nm laser. These results indicate the great potential of WO3 nanowires for applications in fabricating high performance visible photodetectors.

[Comparison of the clinical characteristics between RNA positive and negative patients clinically diagnosed with coronavirus disease 2019].

Objective: To raise awareness about 2019 novel coronavirus pneumonia (NCP) and reduce missed diagnosis rate and misdiagnosis rate by comparing the clinical characteristics between RNA positive and negative patients clinically diagnosed with NCP. Methods: From January 2020 to February 2020, 54 patients who were newly diagnosed with NCP in Wuhan Fourth Hospital were included in this study. RT-PCR method was used to measure the level of 2019-nCov RNA in pharyngeal swab samples of these patients. The patients were divided into RNA positive and negative group, and the differences of clinical, laboratory, and radiological characteristics were compared. Results: There were 31 RNA of 2019-nCov positive cases, and 23 negative cases. Common clinical symptoms of two groups were fever (80.64% vs. 86.96%) , chills (61.29% vs. 52.17%) , cough (80.64% vs. 95.65%) , fatigue (61.30% vs. 56.52%) , chest distress (77.42% vs.73.91%) . Some other symptoms were headache, myalgia, dyspnea, diarrhea, nausea and vomiting. The laboratory and radiological characteristics of two groups mainly were lymphopenia, increased erythrocyte sedimentation rate, increased C-reactive protein, increased lactate dehydrogenase, decreased oxygenation index, normal white blood cell count and bilateral chest CT involvement. There was no statistically significant difference in other clinical characteristics except for dyspnea between two groups. Conclusions: RNA positive and negative NCP patients shared similar clinical symptoms, while RNA positive NCP patients tended to have dyspnea. Therefore, we should improve the understanding of NCP to prevent missed diagnosis and misdiagnosis; In addition, more rapid and accurate NCP diagnostic approaches should be further developed.

Tubular basement membrane immune complex deposition is associated with activity and progression of lupus nephritis: a large multicenter Chinese study.

Tubulointerstitial injury is found frequently in lupus nephritis. Immune complex deposits can occur in the tubular basement membranes (TBMs), although its significance in lupus nephritis patients remains unclear. This study assessed the clinical and prognostic features of lupus nephritis patients with TBM deposits in a large Chinese multicenter cohort. Complete data were collected from 195 patients with renal biopsy-proven lupus nephritis diagnosed in the Peking University First Hospital as the discovery cohort. A total of 102 lupus nephritis patients were enrolled from another four centers as the validation cohort. The status of TBM deposits was retrospectively assessed using electron microscopy, and the associations of the deposits with clinical data, pathological characteristics and renal outcomes were further analyzed. The percentage of positive TBM deposits was nearly 30% in the lupus nephritis patients. Using immuno-gold labeling, we found that 10/10 patients were positive for IgG, 7/10 were C3d positive, 6/10 were C1q positive, and 1/10 were C4d positive. Patients with TBM deposits presented with more active features, including a higher SLEDAI score (SLE Disease Activity Index) ( p < 0.001), higher serum creatinine level ( p = 0.001) and lower serum C3 level ( p < 0.001). These patients also presented with higher scores for most renal pathological indices, including the total activity indices score ( p < 0.001) and total chronicity indices score ( p = 0.001). TBM deposits affected renal outcomes in the univariate Cox hazards regression analysis (HR = 4.2, 95% CI = 1.3-14.3, p = 0.02). In conclusion, TBM deposits were common in lupus nephritis patients and correlated closely with the clinical disease activity and renal outcome.

[Clinical efficacy of anterior decompression and stability reconstruction in patients with cervical hyperextension injury].

Objective: To evaluate the clinical efficacy of anterior decompression and stability reconstruction in patients with cervical hyperextension injury. Methods: Postoperative data from 60 patients with cervical hyperextension injury between April 2009 and December 2016 were analyzed retrospectively; the patients included 50 males and 10 females, aged 21-87 years [average, (57±13) years]. All patients had various degrees of spinal cord injury, and were treated with anterior cervical decompression, fusion, and internal fixation.The preoperative and postoperative neurological function were compared to evaluate the clinical efficacy of the treatment.The t test was applied when preoperative and postoperative data were compared. Results: Of the 60 patients, 5 underwent anterior cervical discectomy and fusion, 26 underwent anterior cervical corpectomy and fusion, and 29 received treatment with the hybrid technique.The average follow-up was (5.1±2.1) years (range, 1.6-9.1 years). The American Spinal Injury Association (ASIA) scores, abbreviated injury scale (AIS grades), and Japanese Orthopedic Association Scores (JOA scores) at the 1 week post operation and final follow-up were significantly better than those obtained preoperatively (all P<0.01). The JOA scores before operation and at the final follow-up was 10(7, 11) and 16(14, 17), respectively.Based on the recovery rate of JOA scores, the rate of cure was 28.3% (17 cases), the rate of significant efficiency was 60.0% (36 cases), the rate of efficiency was 8.3% (5 cases), the rate of inefficiency was 3.3% (2 cases). Among the patients showing cure, 5 were satisfied with the life function, however, the extent of their injury was still of Grade D owing to the incomplete recovery of muscle force.Further, when the 60 patients were divided into A and B groups according to whether they had congenital spinal stenosis or not, respectively, and no significant difference was found in ASIA scores, AIS grades and JOA scores between the groups (all P<0.01). Conclusion: Anterior cervical decompression, fusion and internal fixation is safe and effective for cervical hyperextension injury without continuous-type ossification of the posterior longitudinal ligament.

[Clinical analysis of lower limb thrombosis caused by paraquat poisoning].

Objective: To investigate the causes of peripheral vascular thrombosis in patients with paraquat poisoning. Methods: The patients with paraquat poisoning who were admitted to our department in recent two years were observed to screen out the patients with large vessel thrombosis. The data on toxic exposure history, clinical features, and treatment were collected to analyze the causes of thrombosis in the patients with paraquat poisoning. Results: Three patients had typical lower limb thrombosis. There was one case of right common femoral vein thrombosis, one case of bilateral calf muscle vein thrombosis, and one case of right calf superficial vein thrombosis and right calf muscle vein thrombosis. Conclusions: After paraquat poisoning, the blood is in a hypercoagulable state and prolonged bed rest may increase the risk of thrombosis.

Genome-wide association analysis identifies the genetic basis of fat deposition in the tails of sheep (Ovis aries).

Fat-tailed sheep (Ovis aries) can survive in harsh environments and satisfy human's intake of dietary fat. However, the animals require more feed, which increases the cost of farming. Thus, most farmers currently prefer thin-tailed, short-tailed or docked sheep. To date, the molecular mechanism of the formation of fat tails in sheep has not been completely elucidated. Here, we conducted a genome-wide association study using phenotypes and genotypes (the Ovine Infinium HD SNP BeadChip genotype data) of two breeds of contrasting tail types (78 Small-tailed and 78 Large-tailed Han sheep breeds) to identify functional genes and variants associated with fat deposition. We identified four significantly (rs416433540, rs409848439, rs408118325 and rs402128848) and three approximately associated autosomal SNPs (rs401248376, rs402445895 and rs416201901). Gene annotation indicated that the surrounding genes (CREB1, STEAP4, CTBP1 and RIP140, also known as NRIP1) function in lipid storage or fat cell regulation. Furthermore, through an X-chromosome-wide association analysis, we detected significantly associated SNPs in the OARX: 88-89 Mb region, which could be a strong candidate genomic region for fat deposition in tails of sheep. Our results represent a new genomic resource for sheep genetics and breeding. In addition, the findings provide novel insights into genetic mechanisms of fat deposition in the tail of sheep and other mammals.

A genome-wide association study reveals candidate genes for the supernumerary nipple phenotype in sheep (Ovis aries).

Genome-wide association studies (GWASs) have been widely applied in livestock to identify genes associated with traits of economic interest. Here, we conducted the first GWAS of the supernumerary nipple phenotype in Wadi sheep, a native Chinese sheep breed, based on Ovine Infinium HD SNP BeadChip genotypes in a total of 144 ewes (75 cases with four teats, including two normal and two supernumerary teats, and 69 control cases with two teats). We detected 63 significant SNPs at the chromosome-wise threshold. Additionally, one candidate region (chr1: 170.723-170.734 Mb) was identified by haplotype-based association tests, with one SNP (rs413490006) surrounding functional genes BBX and CD47 on chromosome 1 being commonly identified as significant by the two mentioned analyses. Moreover, Gene Ontology enrichment for the significant SNPs identified by the GWAS analysis was functionally clustered into the categories of receptor activity and synaptic membrane. In addition, pathway mapping revealed four promising pathways (Wnt, oxytocin, MAPK and axon guidance) involved in the development of the supernumerary nipple phenotype. Our results provide novel and important insights into the genetic mechanisms underlying the phenotype of supernumerary nipples in mammals, including humans. These findings may be useful for future breeding and genetics in sheep and other livestock.

[Expression characteristics of epithelial markers in human embryonic stem cells differentiating into keratinocytes].

OBJECTIVE: To differentiate human embryonic stem cells (hESCs) into keratinocytes (K-hESCs) and analyse the expression characteristics of biomarkers of K-hESCs. METHODS: The hESCs of line H9 were seeded on matrigel in mTeSR1 medium. The hESCs were directly differentiated into keratinocytes in epithelial differentiation medium with bone morphogenetic protein 4, retinoic acid and N2 supplement. The karyotype of K-hESCs was analyzed, comparing the gene expression differences of K-hESCs with human gingival epithelial cells (HGECs), human immortalized oral epithelial cells (HIOECs) and HaCaT by Real-time PCR. Molecular characteristics of the cell differentiation were observed throughout the process by immunocytochemical techniques. RESULTS: H9-hESCs were successfully differentiated into the cells that exhibited characteristics of keratinocytes in epithelial differentiation medium. The karyotype of K-hESCs was 46, XX; and the keratinocyte gene p63 expression in K-hESCs was significantly lower than that in HaCaT (P < 0.05), but there was no significant difference of p63 expression in K-hESCs, comparing with that in HGECs and HIOECs (P > 0.05). CONCLUSION: H9-hESCs could be directly differentiated into K-hESCs. The gene expression of K-hESCs was similar to that of epithelial cells in the early stage of monolayer cells differentiation with high proliferative activity.

[A preliminary evaluation and discussion on the value of the medical content of book 'Zhu Bing â ', a medical bamboo compilation excavated from a Han dynasty tomb in Laoguanshan].

The compilation of medical bamboo slips, which are excavated from a Han dynasty tomb in Laoguanshan(Mount. Laoguan), has two manuscripts: 'Zhu Bing Ⅰ(cases study Ⅰ)'and 'Zhu Bing Ⅱ(cases study Ⅱ)'. 'Zhu Bing Ⅰ'mainly discussed the pathogenic factor "wind" and "wind diseases" . It also analyzed the features of pathogenic factor "wind" . It classified the "wind diseases" into 17 kinds from 4 point views, and documented the symptoms of 17 kinds of "wind disease" . It was first time that its most ideas were documented in traditional Chinese medicine literatures.

Endothelium-derived relaxing factor/nitric oxide modulates angiotensin II action in the isolated microperfused rabbit afferent but not efferent arteriole.

It has been reported that sensitivity to angiotensin II (Ang II) is higher in efferent (Ef) than afferent (Af) arterioles (Arts). We tested the hypothesis that this is due to arteriolar differences in the interaction between Ang II and endothelium-derived relaxing factor/nitric oxide (EDNO). Rabbit Af-Arts with glomerulus intact were microperfused in vitro at a constant pressure. Ef-Arts were perfused from the distal end of either the Af-Art (orthograde perfusion) or the Ef-Art (retrograde perfusion) to eliminate influences of the Af-Art or glomerulus, respectively. Ang II did not alter Af-Art luminal diameter until the concentration reached 10(-9) M, which decreased the diameter by 11 +/- 2.6% (n = 11; P < 0.002). In contrast, Ef-Arts became significantly constricted at concentrations as low as 10(-11) M with either perfusion. Surprisingly, the decrease in Ef-Art diameter at 10(-10), 10(-9), and 10(-8) M was significantly greater with retrograde perfusion (44 +/- 6.9%, 70 +/- 5.6%, and 74 +/- 4.1%, respectively; n = 5) than with orthograde perfusion (16 +/- 4.2%, 25 +/- 2.9%, and 35 +/- 3.5%; n = 9). ENDO synthesis inhibition with 10(-4) M nitro-L-arginine methyl ester (L-NAME) decreased the diameter to a greater extent in Af-Arts (22 +/- 3.0%; n = 11) compared to Ef-Arts with either orthograde (9.5 +/- 2.3%; n = 8) or retrograde perfusion (1.2 +/- 2.1%; n = 6). With L-NAME pretreatment, Af-Art constriction induced by 10(-10) M (14 +/- 4.0%, n = 9) and 10(-9) M Ang II (38 +/- 3.9%) was significantly greater compared to nontreated Af-Arts. In contrast, L-NAME pretreatment had no effect on Ang II-induced constriction in Ef-Arts with either perfusion. In conclusion, this study demonstrates higher sensitivity of Ef-Arts to Ang II, particularly with retrograde perfusion. Our results suggest that EDNO significantly modulates the vasoconstrictor action of Ang II in Af-Arts II but not Ef-Arts, contributing to the differential sensitivity to Ang II.

Vasodilator and constrictor actions of platelet-activating factor in the isolated microperfused afferent arteriole of the rabbit kidney. Role of endothelium-derived relaxing factor/nitric oxide and cyclooxygenase products.

It has been suggested that platelet-activating factor (PAF) plays a prominent role in the control of glomerular hemodynamics in various physiological and pathological conditions. We examined the direct effect of PAF on rabbit glomerular afferent arterioles (Af-Arts) microperfused in vitro and tested whether endothelium-derived relaxing factor/nitric oxide (EDNO) and cyclooxygenase products are involved in its actions. In nanomolar concentrations PAF caused dose-dependent constriction of Af-Arts, with the maximum constriction being 34 +/- 10% at 4 x 10(-8) M (n = 9, P < 0.001). The constriction was blunted by cyclooxygenase inhibition (11 +/- 6%, n = 7, P < 0.05) but augmented by EDNO inhibition (76 +/- 14%, n = 8, P < 0.005). To study a possible vasodilator effect of PAF, Af-Arts were preconstricted with norepinephrine and increasing concentrations of PAF added to the lumen. At picomolar concentrations (lower than those that caused constriction), PAF produced dose-dependent vasodilation that was unaffected by cyclooxygenase inhibition but was abolished by EDNO synthesis inhibition. Both PAF-induced constriction and dilation of Af-Arts were blocked by a PAF receptor antagonist. This study demonstrates that PAF has a receptor-mediated biphasic effect on rabbit Af-Arts, dilating them at low concentrations while constricting them at higher concentrations. Our results suggest that PAF's vasodilator action may be due to production of EDNO, while its constrictor action is mediated at least in part through cyclooxygenase products.

Simultaneous determination of two active components in compound aspirin tablets using principal component artificial neural networks (PC-ANNs) on NIR spectroscopy.

A method for simultaneous, non-destructive analysis of aspirin and phenacetin in compound aspirin tablets with different concentrations has been developed by principal component artificial neural networks (PC-ANNs) on near-infrared (NIR) spectroscopy. In PC-ANNs models, the spectra data were first analyzed by principal component analysis (PCA). Then the scores of the principal compounds (PCs) were chosen as input nodes for input layer instead of the spectra data. The artificial neural networks (ANNs) models using the spectra data as input nodes were also established, which were compared with the PC-ANNs models. Four different preprocessing methods (first-derivation, second-derivation, standard normal variate (SNV) and multiplicative scatter correction (MSC)) were applied to NIR conventional spectra. The result shows the first-derivative model of PC-ANNs multivariate calibration has the lowest training errors and predicting errors. The concept of the degree of approximation was introduced and performed as the selective criterion of the optimum network parameters.

The beta 4 subunit cytoplasmic domain mediates the interaction of alpha 6 beta 4 integrin with the cytoskeleton of hemidesmosomes.

The alpha 6 beta 4 integrin is structurally distinct from all the other known integrins because the cytoplasmic domain of beta 4 is unusually large and contains four type III fibronectin-like modules toward its C-terminus. To examine the function of the beta 4 cytoplasmic tail, we have expressed full-length and truncated human beta 4 cDNAs in rat bladder epithelial 804G cells, which form hemidesmosome-like adhesions in vitro. The cDNA encoded wild-type beta 4 subunit associated with endogenous alpha 6 and was recruited at the cell surface within hemidesmosome-like adhesions. A recombinant form of beta 4, lacking almost the entire cytoplasmic domain associated with alpha 6, reached the cell surface but remained diffusely distributed. A beta 4 molecule lacking almost the entire extracellular portion did not associate with alpha 6 but was correctly targeted to the hemidesmosome-like adhesions. Thus, the cytoplasmic portion of beta 4 contains sequences that are required and may be sufficient for the assembly of the alpha 6 beta 4 integrin into hemidesmosomes. To localize these sequences we examined the properties of additional mutant forms of beta 4. A truncated beta 4 subunit, lacking the most C-terminal pair of type III fibronectin homology domains, was incorporated into hemidesmosome-like adhesions, but another recombinant beta 4 molecule, lacking both pairs of type III fibronectin repeats, was not. Finally a recombinant beta 4 molecule, which was created by adjoining the region of the cytoplasmic domain including all type III repeats to the transmembrane segment, was efficiently recruited in hemidesmosome-like adhesions. Taken together these results suggest that the assembly of the alpha 6 beta 4 integrin into hemidesmosomes is mediated by a 303-amino acid region of beta 4 tail that comprises the first pair of type III fibronectin repeats and the segment between the second and third repeats. These data imply a function of a specific segment of the beta 4 cytoplasmic domain in interaction with cytoskeletal components of hemidesmosomes.

[A preliminary discussion on the contents and value of All Diseases(1), the medical bamboo slips unearthed from Han tomb in Laoguanshan of Chengdu].

Among the medical bamboo slips unearthed from Han tomb in Laoguanshan of Chengdu, the Zhu bing (All Diseases) is a monograph to discuss the characteristics of signs and symptoms. Based on the differences of writing styles, diseases involved, expounding methods and writing rules, the book is divided, by the research team, into 2 parts: All Diseases(1) and All Diseases(2). All Diseases(1) includes over 130 slips, 2 000 characters with totally more than 100 disease names, containing multiple clinical disciplines. The elaborated classification, varied naming methods, grasping the symptom characteristics guided by the four diagnostic approaches, paying attention to the comparison of similar diseases, and dealing with the prognosis and healthcare in this part reflect the holism of correspondence between human body and natural environment, and syndrome differentiation thought of combining disease with symptoms and signs, revealing its academic significance.

[A preliminary discussion on the contents and value of Shi er mai (Twelve Channels) and Bie mai (Allo-channels), the medical bamboo slips unearthed from Han tomb in Laoguanshan of Chengdu].

The 52 medical bamboo slips entitled Si er ma (Twelve Channels) and Bie mai (Allo-channels) , unearthed from Han tomb in Laoguanshan, are the exclusive medical books concerning channels. Shi er mai is the first work bearing the term " heart dominating channel" , and the running pathways and manifestations of 12 channels. This book could be the major original sources of the 12-channel theories described in the Chapter of Jing mai (Channels) of Ling shu (Miraculous Pivot) . While Bie mai suggests the coexistence of the multi-channel systems at the time when the book was compiled and, together with the 12 "channel" , and 3 "branched collaterals" , these 2 books represent the original condition before the establishment of the mode of channel circulation in Ling shu, and is significant to explore the nature and origin of channel.