Study on the relationship between intestinal flora analysis and CD4 +T lymphocyte subsets in patients with systemic lupus erythematosus / 中华风湿病学杂志

Objective:To explore the characteristics of intestinal microbiota in patients with systemic lupus erythematosus (SLE), and further explore the relationship between microbiota and CD4 +T lymphocyte subsets and disease activity. Methods:Fecal samples were collected from 96 patients with SLE, and 96 sex- and age-matched healthy controls (HCs). The gut microbiota were investigated via 16s rRNA sequencing. Flow cytometry was used to detect peripheral CD4 +T lymphocyte subsets of Th1, Th2, Th17 and Treg cells. Indicators of disease activity such as erythrocyte sedimentation rate (ESR), C-reactive protein (CRP), complement C3 and C4, Systemic lupus erythematosus disease activity index(SLEDAI) for each patient were recorded. Differential abundance analysis was carried out using the edgeR algorithm. The Wilcoxon rank-sum test was used to compare alpha diversity indices, bacterial abundances, and the F/B ratio between groups. R (version 4.0.1) was used for comparative statistics, and Pearson′s correlation analysis was used to assess the correlations between the relative abundances of bacterial genera and serum levels of ESR, CRP, C3 and C4 in the samples. Results:The alpha estimators of richness (ACE and Chao 1) were significantly reduced in SLE feces samples compared with those of HCs ( P<0.01). Bacterial diversity estimators, including the Shannon ( P<0.01) and Simpson′s ( P<0.01) indices, were also significantly lower in SLE. Significant differences in gut microbiota composition between SLE and HCs were found using the edgeR algorithm. Compared with HC, 24 species of bacteria were significantly different in SLE patients at the genus level ( P<0.05). Moreover, there was a significant positive correlation between CRP and Coprococcus ( r=0.30, P=0.014), C4 and Corynebacterium ( r=0.31, P=0.013) and Faecalibacterium( r=0.25, P=0.048), Hemoglobin and Morganella( r=0.41, P=0.001), as well as SLIDA and Corynebacterium( r=0.25, P=0.047). In terms of lymphocyte subsets, there was significant positive correlation between B cells, Treg cells and Eubacterium eligens group, as well as CD8 +T, CD4 +T, NK cells and Corynebacterium. In additional, Th1 was positively correlated with Shigella Escherichia coli ( r=0.52, P=0.008), and Th2 was positively correlated with Dielma ( r=0.51, P<0.001). Conclusion:The abundance and diversity of intestinal flora in SLE patients were significantly reduced, and the differentially expressed bacteria were closely related to the CD4 +T lymphocyte subsets and disease activity indicators of patients.

Jiawei Xiaoyao San in treatment of anxiety disorder and anxiety: A review / 中草药·英文版

Jiawei Xiaoyao San (JWXYS) has shown excellent clinical efficacy in anxiety disorder, but has not yet attracted widespread attention. The animal experiments, clinical trials and mechanism studies of JWXYS were reviewed in this article, which may provide a reference for developing new anxiolytic drugs based on this prescription. The literature was searched in PubMed and CNKI and the documents written in English or with English abstracts were selected. JWXYS could reduce the anxiety symptoms of patients alone and reduce the adverse reactions when it is used in combination with other drugs in the clinic. In preclinical studies, JWXYS also showed therapeutic effects in reducing anxiety-like behavior. The mechanisms may include improving the hypothalamic-pituitaryadrenal (HPA) axis and hormone disorders, increasing neurotransmitter content, neurogenesis, and regulating the synthesis of related enzymes. This article shows that JWXYS could effectively treat anxiety disorders by regulating the central nervous system. In the future, with the participation of more researchers, it is expected to develop innovative drugs for the treatment of anxiety disorders based on JWXYS.

CD301b+ macrophage: the new booster for activating bone regeneration in periodontitis treatment / 国际口腔科学杂志·英文版

Periodontal bone regeneration is a major challenge in the treatment of periodontitis. Currently the main obstacle is the difficulty of restoring the regenerative vitality of periodontal osteoblast lineages suppressed by inflammation, via conventional treatment. CD301b+ macrophages were recently identified as a subpopulation that is characteristic of a regenerative environment, but their role in periodontal bone repair has not been reported. The current study indicates that CD301b+ macrophages may be a constituent component of periodontal bone repair, and that they are devoted to bone formation in the resolving phase of periodontitis. Transcriptome sequencing suggested that CD301b+ macrophages could positively regulate osteogenesis-related processes. In vitro, CD301b+ macrophages could be induced by interleukin 4 (IL-4) unless proinflammatory cytokines such as interleukin 1β (IL-1β) and tumor necrosis factor α (TNF-α) were present. Mechanistically, CD301b+ macrophages promoted osteoblast differentiation via insulin-like growth factor 1 (IGF-1)/thymoma viral proto-oncogene 1 (Akt)/mammalian target of rapamycin (mTOR) signaling. An osteogenic inducible nano-capsule (OINC) consisting of a gold nanocage loaded with IL-4 as the "core" and mouse neutrophil membrane as the "shell" was designed. When injected into periodontal tissue, OINCs first absorbed proinflammatory cytokines in inflamed periodontal tissue, then released IL-4 controlled by far-red irradiation. These events collectively promoted CD301b+ macrophage enrichment, which further boosted periodontal bone regeneration. The current study highlights the osteoinductive role of CD301b+ macrophages, and suggests a CD301b+ macrophage-targeted induction strategy based on biomimetic nano-capsules for improved therapeutic efficacy, which may also provide a potential therapeutic target and strategy for other inflammatory bone diseases.

Identification of an LDLR variant in a Chinese familial hypercholesterolemia and its relation to ROS/NLRP3-Mediated pyroptosis in hepatic cells / 老年心脏病学杂志（英文版）

BACKGROUND@#Familial hypercholesterolemia (FH) is a common autosomal dominant hereditary disease. Its early diagnosis and intervention significantly improve the patient's quality of life. However, there are few types of research on the FH pathogenic genes in China.@*METHODS@#In this study, we recruited a family diagnosed with FH and used whole exome sequencing (WES) to analyze the proband variants. Intracellular cholesterol level, reactive oxygen species (ROS) level, and the expression of pyroptosis-related genes were detected after overexpression of wild-type or variant LDLR in L02 cells.@*RESULTS@#A heterozygous missense variant predicted to be deleterious to LDLR (c.1879G > A, p.Ala627Thr) was identified in the proband. Mechanistically, intracellular cholesterol level, ROS level, and the expression of pyroptosis-related genes, nucleotide-binding oligomerization domain-like receptor family protein 3 (NLRP3) inflammasome and components (caspase 1, apoptosis-associated speck-like protein containing a caspase recruitment domain (ASC) and NLRP3), gasdermin D (GSDMD), interleukin (IL) -18, IL-1β was elevated in the variant LDLR group, which was attenuated by inhibition of ROS.@*CONCLUSIONS@#FH is associated with a variant (c.1879G>A, p.Ala627Thr) in the LDLR gene. Regarding the mechanism, the ROS/NLRP3-mediated pyroptosis in hepatic cells may contribute to the pathogenesis of the LDLR variant.

The role of ROS/TXNIP/NLRP3 pathway in the skin injury of trichloroethylene sensitized mice / 中华劳动卫生职业病杂志

Objective: To explore the mechanism of reactive oxygen species/thioredoxin-interacting protein/nucleotide-binding oligomerization domain-like receptor 3 (ROS/TXNIP/NLRP3) pathway in the skin injury of trichloroethylene (TCE) sensitized mice. Methods: In August 2020, 40 female BALB/c mice were randomly divided into control group (n=5) , solvent control group (n=5) , TCE treatment group (n=15) and TCE+(2-(2, 2, 6, 6-Tetrameyhylpiperidin-1-oxyl-4-ylamino)-2-oxoethyl) triphenylphosphonium chloride (Mito TEMPO) treatment group (n=15) . The TCE sensitization model was established. Mice in the TCE treatment group and TCE+Mito TEMPO treatment group were divided into the sensitized positive group and the sensitized negative group according to the skin erythema and edema reactions on the back of the mice 24 h after the last stimulation. The mice were sacrificed 72 h after the last stimulation, the back skin of the mice was taken, and the skin lesions were observed. Immunohistochemistry (IHC) was used to detect the expression level of NLRP3, and the Western Blot was performed to detect the expression levels of NLRP3, apoptosis-associated speck-like protein containing a CARD (ASC) , cysteinyl aspartate specific proteinase 1 (Caspase 1) , Interleukin-1β (IL-1β) and TXNIP proteins in the skin of the mice, the reactive oxygen species (ROS) kit was used to detect the level of intracellular ROS in the back skin tissue. Results: The sensitization rates of TCE treatment group and TCE+Mito TEMPO treatment group were 40.0% (6/15) and 33.3% (5/15) , respectively, and there was no significant difference between the two groups (P>0.05) . The back skin of the mice in the TCE sensitized positive group was thickened and infiltrated by a large number of inflammatory cells. The number of mitochondria in the epidermis cells was significantly reduced, the mitochondrial crest disappeared and vacuolar degeneration occurred. TCE+Mito TEMPO sensitized positive group had less damage, more mitochondria and relatively normal cell structure. Compared with the solvent control group and corresponding sensitized negative groups, the expression levels of NLRP3, ASC, Caspase 1, IL-1β, TXNIP proteins and the content of ROS in the TCE sensitized positive group and TCE+Mito TEMPO sensitized positive group were significantly increased (P<0.05) . Compared with TCE sensitized positive group, the expression levels of NLRP3, ASC, Caspase 1, IL-1β, TXNIP proteins and the content of ROS in the TCE+Mito TEMPO sensitized positive group were significantly decreased (P<0.05) . Conclusion: ROS/TXNIP/NLRP3 pathway was activated and then encouraged the release of IL-1β, finally aggravated the TCE-induced skin injury.

Participation of clinical pharmacists in pharmaceutical care for a patient with Mycobacterium abscess infection after the resection of intervertebral schwannoma / 中国药房

OBJECTIVE To provide ideas for pharmaceutical care of Mycobacterium abscess infection. METHODS The diagnosis and treatment plan ，about a patient with M. abscess infection after resection of intervertebral neurilemmoma participated by clinical pharmacists ，was analyzed . According to the results of bacterial culture and drug sensitivity test of wound secretion ，the clinical pharmacist timely dealed with the adverse drug reactions during the treatment . For the tinnitus caused by the patient taking amikacin（0.4 g，intravenous drip ，q12 h），it was suggested to stop amikacin and replace it with linezolid ；however，the patient suffered from knee pain after taking linezolid . The clinical pharmacist once again suggested stopping linezolid ，continuing to use amikacin，and adjusting the dose to 0.8 g，intravenous drip ，q24 h；at the same time ，the patients were given medication guidance and ordered to have regular follow -up after discharge . RESULTS The physicians adopted the suggestions of clinical pharmacists ， and the patient did not suffer from tinnitus ，knee pain and other discomfort ，and the liver and kidney functions were normal . CONCLUSIONS Clinical pharmacists timely assist clinicians to adjust the medication plan ，which improves the effectiveness and safety of patients ’medication.

Spatio-temporal distribution characteristics of Oncomelania hupensis in Anhui Province from 2011 to 2020 / 中国血吸虫病防治杂志

OBJECTIVE@#To analyze the spatial-temporal distribution characteristics of Oncomelania hupensis snails in Anhui Province from 2011 to 2020, to provide insights into precision control of O. hupensis snails in Anhui Province.@*METHODS@#O. hupensis snail distribution data were collected in Anhui Province from 2011 to 2020 and descriptively analyzed, including actual area of snail habitats, area of emerging snail habitats and area of Schistosoma japonicum-infected snails. The actual area of snail habitats and area of emerging snail habitats were subjected to spatial autocorrelation analysis, hotspot analysis, standard deviation ellipse analysis and space-time scanning analysis, and the clusters of snail distribution and settings at high risk of snail spread were identified in Anhui Province from 2011 to 2020.@*RESULTS@#The actual area of snail habitats gradually decreased in Anhui Province from 2011 to 2020. The actual area of snail habitats were 26 238.85 hm2 in Anhui Province in 2020, which were mainly distributed in marshland and lake regions. There was a large fluctuation in the area of emerging snail habitats in Anhui Province during the period from 2011 to 2020, with the largest area seen in 2016 (1 287.65 hm2), and 1.96 hm2 emerging infected snail habitats were detected in Guichi District, Chizhou City in 2020. Spatial autocorrelation and hotspot analyses showed spatial clusters in the distribution of actual areas of snail habitats in Anhui Province from 2011 to 2020 (Z = 3.00 to 3.43, all P values < 0.01), and the hotspots were mainly concentrated in the marshland and lake regions and distributed along the south side of the Yangtze River, while the cold spots were mainly concentrated in the mountainous regions of southern Anhui Province. There were no overall spatial clusters in the distribution of areas of emerging snail habitats (Z = -2.20 to 1.71, all P values > 0.05), and a scattered distribution was found in local regions. Standard deviation ellipse analysis showed relatively stable distributions of the actual areas of snail habitats in Anhui Province from 2011 to 2020, which was consistent with the flow direction of the Yangtze River, and the focus of the distribution of areas of emerging snail habitats shifted from the lower reaches to upper reaches of Anhui section of the Yangtze River. Space-time scanning analysis identified two high-value clusters in the distribution of actual areas of snail habitats in lower and middle reaches of Anhui section of the Yangtze River from 2011 to 2020, and two high-value clusters in the distribution of areas of emerging snail habitats were identified in mountainous and hilly regions.@*CONCLUSIONS@#There were spatial clusters in the distribution of O. hupensis snails in Anhui Province from 2011 to 2020, which appeared a tendency of aggregation towards the south side and upper reaches of the Yangtze River; however, the spread of O. hupensis snails could not be neglected in mountainous and hilly regions. Monitoring of emerging snail habitats should be reinforced in mountainous and hilly regions and along the Yangtze River basin.

Calcifying fibrous tumor of the stomach: report of nine cases and review of literature / 肿瘤研究与临床

Objective:To investigate the clinicopathological features and pathological diagnosis methods of calcifying fibrous tumor (CFT) of the stomach.Methods:The clinicopathological data of 9 patients with gastric CFT in the Second Hospital of Hebei Medical University from August 2015 to May 2020 were retrospectively analyzed, immunohistochemistry was used to detect the immunophenotypic characteristics of gastric CFT, and the relevant literature was reviewed.Results:Among 9 patients with gastric CFT, 1 case was male and 8 cases were female, with a median age of 53 years old (18-63 years old). There were 7 cases occurred in the body of the stomach, while 2 cases in the fundus. All cases were single solid nodule with the cut surface gray-white or gray-red in colour and hard and tough in texture. Maximum diameter of tumor ranged from 0.6 cm to 1.5 cm. Observation under the microscope showed the spindle-shaped tumor cells were sparsely distributed in a large amount of collagenized stroma with scattered lymphocytes and plasma cells infiltration. Calcification or gravel formation was detected in all cases. The tumor cells in 9 cases expressed Vimentin, 3 cases expressed CD34, none of them expressed PDGFRA, SMA, SMMS-1, Desmin, ALK, DOG-1, CD117, S-100, β-catenin, CKpan and calponin; Ki-67 positive index was 1%-3%.Conclusions:Gastric CFT is more common in middle-aged and elderly people, which is characterized by gastric body with smaller volume. The diagnosis of gastric CFT mainly depends on pathological morphology and immunohistochemical examination. It needs to be differentiated from a variety of spindle cell tumors more commonly found in the stomach.

A predictive analysis of influencing factors with quality of life among patients with gout: based on Wilson-Cleary model / 中国实用护理杂志

Objective:To explore the impact of pain, functional disability, self-efficacy and social support on health-related quality of life (HRQOL) in gout patients.Methods:From November 2019 to January 2020, a total of 218 patients with gout were investigated using the general information questionnaire, Visual Analogue Scale, Health Assessment Questionnaire Disability Index, General Self-Efficacy Scale, Perceived Social Support Scale, and Gout Impact Scale. The structural equation model was established by AMOS 24.0 for parth analysis, and the mechanism of pain dysfuction, self-efficacy and social support affecting the quality of life in gout patients was tested.Results:The total score of Gout Impact Scale, pain, functional disability, self-efficacy and social support respectively was 59.94±18.39, 6.00±2.76, 0.25 0, 0.88, 23.39±6.40 and 62.92±8.24. Pain directly influenced HRQOL ( β=-0.293, P<0.01), and indirectly influenced HRQOL ( β=-0.039, P<0.05). Functional disability directly influenced HRQOL ( β=-0.244, P<0.01). Self-efficacy directly influenced HRQOL ( β=0.182, P<0.01), and indirectly influenced HRQOL ( β=0.202, P<0.01) through pain and functional disability. Social support indirectly influenced HRQOL ( β=0.278, P<0.01) through pain, functional disability and self-efficacy. Conclusions:HRQOL of patients among gout is affected by several factors, mainly affected by pain, functional disability and self-efficacy; and there are interactions among them. Targeted interventions should be strengthened to relieve pain, prevent or slow down the progress of physical disability, enhance self-efficacy and social support to improve HRQOL.

Progress of researches on the application of risk assessment models in assessment of the schistosomiasis transmission risk / 中国血吸虫病防治杂志

With the deepening of the schistosomiasis research, risk assessment models have been widely used in schistosomiasis research and control. This paper reviews the theoretical basis and applications of common schistosomiasis risk assessment models and the Bayesian model, so as to provide insights into national schistosomiasis elimination program in China.

Clinical observation on wheat grain moxibustion for chronic superficial gastritis of spleen-stomach deficiency and cold / 中国针灸

OBJECTIVE@#To compare the clinical effect of wheat grain moxibustion and warming acupuncture on chronic superficial gastritis of spleen-stomach deficiency and cold.@*METHODS@#A total of 200 patients with chronic superficial gastritis of spleen-stomach deficiency and cold were randomly divided into a wheat grain moxibustion group (100 cases, 13 cases dropped off ) and a warming acupuncture group (100 cases, 16 cases dropped off ). The acupoints of Zhongwan (CV 12), Tianshu (ST 25), Qihai (CV 6), Liangqiu (ST 34) and Zusanli (ST 36) were selected in both groups. The patients in the wheat grain moxibustion group were treated with wheat grain moxibustion, and 5 cone were given on every acupoint each time. Moxibustion was performed after acupuncture in the warming acupuncture group, and 3 cone each acupoint. Both groups were treated every other day for 15 times. The scores of clinical symptoms and signs, scores of physical component summary (PCS) and mental component summary (MCS) of quality of life in the two groups were recorded before and after treatment and during follow-up 1 month after treatment, and the clinical effects of the two groups were evaluated after treatment and during follow-up.@*RESULTS@#After treatment and during follow-up, the scores of clinical symptoms and signs of the two groups were lower than before treatment (@*CONCLUSION@#The wheat grain moxibustion can effectively improve the clinical symptoms, signs and quality of life in patients with chronic superficial gastritis of spleen-stomach deficiency and cold, and its long-term effect is better than warming acupuncture.

Abnormal metabolism of gut microbiota reveals the possible molecular mechanism of nephropathy induced by hyperuricemia

The progression of hyperuricemia disease is often accompanied by damage to renal function. However, there are few studies on hyperuricemia nephropathy, especially its association with intestinal flora. This study combines metabolomics and gut microbiota diversity analysis to explore metabolic changes using a rat model as well as the changes in intestinal flora composition. The results showed that amino acid metabolism was disturbed with serine, glutamate and glutamine being downregulated whilst glycine, hydroxyproline and alanine being upregulated. The combined glycine, serine and glutamate could predict hyperuricemia nephropathy with an area under the curve of 1.00. Imbalanced intestinal flora was also observed. , , , , and other conditional pathogens increased significantly in the model group, while and , the short-chain fatty acid producing bacteria, declined greatly. At phylum, family and genus levels, disordered nitrogen circulation in gut microbiota was detected. In the model group, the uric acid decomposition pathway was enhanced with reinforced urea liver-intestine circulation. The results implied that the intestinal flora play a vital role in the pathogenesis of hyperuricemia nephropathy. Hence, modulation of gut microbiota or targeting at metabolic enzymes, , urease, could assist the treatment and prevention of this disease.

Clinical phenotype and genetic analysis of three pedigrees with 17q12 microdeletion syndrome / 中华医学遗传学杂志

OBJECTIVE@#To explore the genetic etiology of three pedigrees with a gestational history of fetal renal anomalies.@*METHODS@#Peripheral venous blood or skin samples were derived from the probands of the three pedigrees. Copy number variation sequencing (CNV-seq) was applied to detect alterations of genome CNVs.@*RESULTS@#The patient from pedigree 1 and the fetuses from pedigrees 2 and 3 all carried a heterozygous 17q12 deletion, with the size ranging from 1.4 Mb to 1.48 Mb encompassing the HNF1B gene.@*CONCLUSION@#The diagnosis of 17q12 microdeletion may be difficult during fetal period for its variable phenotypes. Alterations of chromosomal copy numbers need to be excluded in such patients.

Valproic acid concentration and biochemical indexes in epilepsy outpatients during monotherapy or combination therapy / 中南大学学报(医学版)

OBJECTIVES@#Due to the narrow therapeutic window of valproic acid (VPA), grievous adverse reactions such as hepatotoxicity, nephrotoxicity may occur in patients with epilepsy for a long time. This study aimed to explore the effect of VPA concentration on biochemical and routine blood test related to liver, renal, and hematology in epileptic outpatients treated with VPA alone or combined with other antiepileptic drugs.@*METHODS@#A total of 3 194 Chinese epileptic outpatients from Xiangya Hospital, were analyzed in a crude analysis after stratifying through dosage regimens. The plasma VPA concentration was detected by gas chromatography method and then standardized through dosage and body weight. Ten biochemical indexes related to liver, renal, and hematology were evaluated.@*RESULTS@#Of all patients, blood urea nitrogen (BUN), serum creatinine (SCr) level, and erythrocyte count (RBC) showed positive correlations with standardized VPA concentration (=0.494, =0.157, =0.596, respectively), while platelet specific volume (PCT) and blood platelet (PLT) showed negative correlations with standardized VPA concentration (=-5.500, =-0.086, respectively). After stratifying through dosage regimens, significantly positive associations between SCr and standardized VPA concentration were found in the juvenile patients from the monotherapy group and combination therapy group (=1.800, =0.352, respectively). In addition, PLT and leukocyte count (WBC) in the juvenile patients from the combination therapy group were negatively correlated with standardized VPA concentration (=-1.463, =-0.079, respectively), while RBC showed a positive association with standardized VPA concentration in the juvenile patients from the monotherapy group (=0.068).@*CONCLUSIONS@#SCr level is significantly associated with plasma VPA concentration. Drug combination and age are important factors leading to hematological disorders. The finding provides potential theoretical guidance for the rational and safe clinical use of VPA.

Ultrasonographic manifestation and genetic analysis of a fetus with nephronophthisis type 2 / 中华医学遗传学杂志

OBJECTIVE@#To carry out genetic analysis for a family with a fetus manifesting bilateral polycystic renal dysplasia and oligohydramnios at 16 gestational week and a previous history for fetal renal anomaly.@*METHODS@#Ultrasound scan was carried out to detect the morphological changes. Following genetic counselling, the parents had decided to terminate the pregnancy. Fetal kidneys were subjected to histological examination. Target capture and next generation sequencing (NGS) was applied to the abortus to detect potential variants. The results were verified by Sanger sequencing.@*RESULTS@#Histological examination of fetal kidneys revealed cystic changes without cortex, medulla or normal renal structure. NGS has identified a heterozygous c.100+1G>A variant and deletion of exon 3 of the INVS gene, which were respectively inherited from the mother and father.@*CONCLUSION@#Through NGS and Sanger sequencing, the fetus was diagnosed with type II nephronophthisis (NPHP2). Above result can provide guidance for further pregnancy and enforce understanding of clinical features and genetic etiologies for NPHP.

An investigation into the mechanisms underlying the regulatory effect of the E2F6 transcription factor on proliferation and metastasis of malignant melanoma cells through β-catenin signaling pathway / 中华皮肤科杂志

Objective:To determine the expression of the E2F6 transcription factor in human malignant melanoma tissues and cell lines, and to evaluate the effect of E2F6 on proliferation, migration and invasion of a malignant melanoma cell line A375.Methods:Frozen tissues and paraffin-embedded tissue sections were collected from 50 cases of cutaneous malignant melanoma and 30 cases of pigmented nevus in Department of Dermatology, Chongqing Traditional Chinese Medicine Hospital from January 2012 to December 2017. Quantitative reverse transcription-PCR (qRT-PCR) was performed to determine the mRNA expression of E2F6 in the malignant melanoma and pigmented nevus tissues, as well as in 7 malignant melanoma cell lines (HM, A375, WM451, WM35, SK-MEL-1, Hs-695T and MDA-MB-435s) and pigmented nevus cells, and immunohistochemical study and Western blot analysis were conducted to determine the protein expression of E2F6 and β-catenin in the malignant melanoma tissues. An E2F6-inhibiting plasmid and a control plasmid were separately transfected into A375 cells by using a liposome-mediated transfection method, and the E2F6 gene-knockdown efficiency was verified by qRT-PCR and Western blot analysis. Cell counting kit-8 (CCK8) assay, soft-agar plate cloning assay, Transwell migration and invasion assays and 3D cell culture assay were conducted to evaluate the effect of E2F6 gene knockdown on the proliferation, migration and invasion of A375 cells, flow cytometry was performed to detect the cell cycle and apoptosis rate, and Western blot analysis was conducted to determine the protein expression of total β-catenin, activated β-catenin, c-Myc and cyclin D1. The comparison between two groups was carried out by t test, the comparison among several groups by one-way analysis of variance, and multiple comparisons by least significant difference t test; Pearson correlation coefficient was used to analyze the correlation between E2F6 and β-catenin expression in cutaneous malignant melanoma. Results:The E2F6 mRNA expression was significantly higher in the 7 malignant melanoma cell lines than in the pigmented nevus cells (all P < 0.001). qRT-PCR showed that the relative mRNA expression of E2F6 was significantly higher in the cutaneous malignant melanoma tissues (0.000 55 ± 0.000 17) than in the pigmented nevus tissues (0.000 18 ± 0.000 09, t = 3.22, P < 0.001). Both the immunohistochemical study and Western blot analysis showed significantly increased E2F6 protein expression, but decreased β-catenin protein expression in the cutaneous malignant melanoma tissues compared with the pigmented nevus tissues (all P < 0.001). Correlation analysis showed that E2F6 protein expression was negatively correlated with β-catenin expression in the malignant melanoma tissues (immunohistochemical study: r = -0.56, Western blot analysis: r = -0.63, both P < 0.01). After knockdown of the E2F6 gene in A375 cells, the mRNA and protein expression of E2F6 was significantly lower in the E2F6 inhibition group than in the control group ( t = 3.38, 2.76 respectively, both P < 0.001). CCK8 assay showed that the cellular proliferative ability was significantly lower in the E2F6 inhibition group than in the control group ( t = 4.58, P < 0.01) 48 hours after transfection; soft-agar plate cloning assay showed that the colony-formation ratio was significantly lower in the E2F6 inhibition group than in the control group ( t = 2.26, P < 0.001) ; Transwell migration and invasion assays showed that the number of cells crossing the chamber was significantly lower in the E2F6 inhibition group (165 ± 23, 96 ± 11 respectively) than in the control group (376 ± 22, 315 ± 31, t = 3.14, 2.12, respectively, both P < 0.01) ; 3D cell culture assay showed that the cell morphology markedly changed, and the invasive pseudopodia disappeared in the E2F6 inhibition group. Flow cytometry revealed that the proportion of cells at G0-G1 phase and apoptosis rate were significantly higher in the E2F6 inhibition group than in the control group (both P < 0.001). Western blot analysis showed significantly decreased protein expression of β-catenin, activated β-catenin and its downstream target proteins c-Myc and cyclin D1, but significantly increased protein expression of P21 in the E2F6 inhibition group compared with the control group (all P < 0.001) ; additionally, the E2F6 inhibition group showed significantly decreased protein expression of epithelial-mesenchymal transition-related molecules vimentin and N-cadherin, but significantly increased expression of E-cadherin compared with the control group (all P < 0.001) . Conclusions:The E2F6 transcription factor is highly expressed in malignant melanoma. Knockdown of the E2F6 gene in A375 cells can inhibit cell proliferation, migration and invasion by antagonizing the β-catenin signaling pathway.

Accuracy and clinical application of a REF-XT01 uric acid meter / 中华内分泌代谢杂志

Objective:This study aimed to evaluate the accuracy of a portable REF-XT01 uric acid meter in measuring blood uric acid concentration, and to determine whether the results of the uric acid meter could be used to guide the adjustment of uric acid-lowering drugs.Methods:1 551 subjects were enrolled from the Gout Clinical Medical Center of the Affiliated Hospital of Qingdao University. The fasting venous blood was collected and the serum uric acid was measured by an automatic biochemical analyzer. Meanwhile, the capillary blood uric acid was measured by fingertip puncture using the REF-XT01 uric acid meter. Linear regression, intra-group correlation coefficient(ICC), and Bland-Altman plots were used to analyze the uric acid concentration correlation between the biochemical analyzer(sUA BA)and the uric acid meter(sUA UM). The receiver operating characteristic curve(ROC curve)was conducted to evaluate whether sUA UM can be used as a reference for the gout patients to take uric acid-lowering drugs. Results:The regression analysis showed correlation between sUA BA and sUA UM, with the regression formula Y=0.875X+ 39.525( r=0.84, P<0.01)and the ICC was 0.829(95% CI 0.814-0.844, P<0.01). The Bland-Altman diagram showed a good consistence(the absolute deviation was-143.4-114.5 μmol/L, mean deviation was -14.4 μmol/L)between sUA BA and sUA UM. The sensitivity was 96.61%, specificity was 48.81%, and the area under the ROC curve(AUC)was 0.926( P<0.01)when 300 μmol/L was defined as the detection threshold of the uric acid meter, the sensitivity was 90.98%, specificity was 66.78%, and the area under the ROC curve(AUC)was 0.914( P<0.01)when 360 μmol/L was defined as the detection threshold of the uric acid meter. Conclusion:REF-XT01 uric acid meter is applicable for the adjustment of uric acid-lowering drugs for the gout patients, because of its high accuracy for the detection of uric acid.

Plunging ranula presenting as a giant retroauricular mass: A case report / 北京大学学报(医学版)

Plunging ranula is rare and manifests as the submandibular or submental cystic mass, without intra-oral abnormality. It usually causes misdiagnosis and malpractice. This article reported a case of plunging ranula that appeared as a massive mass located behind the left ear. A 6-year-old child presented with recurrent left retroauricular swelling over six months without obvious inducement. The mass showed repeatedly swelling and persistent skin ulcers. Physical examinations described a fluctuant, nontender mass behind the left ear whose size was about 5 cm×5 cm×3 cm. The skin upon the surface of the mass was thin and red, and overflowed yellow slimelike contents. Computed tomography (CT) showed an extensive cystic lesion in the left neck. After local incision and drainage the mass was shrink. With the primary clinical diagnosis of branchial cyst, the patient underwent mass resection through postauricular incision. During the operation, the fistula was traced to the area around the sublingual gland, and the postoperative pathology report demonstrated cystic spaces occurring in soft tissue without lymphoid tissue. One month postoperatively, the patient presented the "egg-white", wire-drawing transparent viscous fluid outflowing from the left external auditory canal, indicating that the fluid in the external auditory canal originated from the sublingual gland and the disease was the plunging ranula presenting as a giant left retroauricular mass. We readmitted the patient to the hospital and the ipsilateral sublingual gland was completely removed in the mouth under general anesthesia. No clinical evidence of recurrence was found at the end of 9 months follow-up. So far, there is no literature reporting plunging ranula behind ear. This case report describes the clinical manifestation, diagnosis, and differential diagnosis and treatment of a case of plunging ranula, and reveals that, other than the submandibular or submental, plunging ranula also could be found in retroauricular region. The plunging ranula is difficult to fully confirm through a clinical character, and usually easy to be misdiagnosed as other tumors, like lymphatic malformation, venous malformations, etc. Radiographic examination and/or puncture fluid observation are required for identification. "Egg-white", wire-drawing mucous secretion is the most important characteristic in clinical diagnosis of sublingual gland cyst. However, if sublingual gland resection, i.e. removal of the source of cyst fluid is performed, a good prognosis can be achieved.

Chinese medicine Yu-Ping-Feng-San attenuates allergic inflammation by regulating epithelial derived pro-allergic cytokines / 中国天然药物

This study aimed to investigate the mechanisms of Yu-Ping-Feng-San (YPFS) on attenuating allergic inflammation in the initial stage of atopic dermatitis (AD). AD mouse model was established with fluorescein isothiocyanate (FITC) sensitization and elicitation. Epithelial barrier structure was observed with transmission electron microscope. The populations of dendritic cells (DCs) and group 2 innate lymphoid cells (ILC2s) were detected by flow cytometry. Human immortalized keratinocyte (HaCaT) cells were stimulated with Poly(I:C)/TNF-α in vitro to assessthymic stromal lymphopoietin (TSLP), interleukin (IL)-33 and nuclear factor-κB (NF-κB) levels or expressions by immunofluorescence, enzyme linked immunosorbent assay (ELISA) and western blot. In the initial stage of AD, ear swelling and infiltration of inflammatory cells in ear tissues were markedly attenuated with YPFS treatments. The damaged structures of ear epithelium and the increased levels of Th2-cytokines induced by FITC were significantly rescued in YPFS-treated mice. The production of pro-allergic cytokines, TSLP and IL-33, as well as the cell populations of their target cells DCs and ILC2s were decreased in AD model, respectively. Likewise, the levels of TSLP and IL-33 in Poly(I:C)/TNF-α-stimulated HaCaT cells showed the same results. Lower levels of p-NF-κB were detected with YPFS treatment, and the expressions of TSLP and IL-33 could be further decreased with inhibiting of NF-κB. Therefore, YPFS attenuates allergic inflammation in the initial stage of AD probably through regulating NF-κB-TSLP/IL-33 pathway, which may provide a novel effective target for the prevention and treatment of allergic diseases.

Phenotype and genetic analysis of three patients with PKHD1 associated autosomal recessive polycystic kidney disease at childhood, teenage and advanced age / 中华医学遗传学杂志

Objective@#The phenotype and genetics of three patients with autosomal recessive polycystic kidney disease (ARPKD) at childhood, teenage and advanced age were analyzed.@*Methods@#Next generation sequencing (NGS) was applied to all the probands. PCR and Sanger sequencing were used to verify the suspicious gene variants screened by NGS in the probands and their family members, and one of the family got prenatal diagnosis.@*Results@#Through NGS, PCR and Sanger sequencing, the 5-yr proband in pedigree 1 was shown to carry compound heterozygous variants of c. 5935G>A(p.G1979R) and c. 5428G>T(p.E1810X) of PKHD1, originated from his parents; In pedigree 2, the 17-ys proband was detected with c. 5512T>C(p.Y1838H) and c. 5935G>A(p.G1979R) variants of PKHD1 orginated from her parents, and her mother also got prenatal diagnosis during the second trimester; In pedigree 3, the 70-ys female proband was found with variants c. 11314C>T (p.R3772X) and c. 3860T>G (p.V1287G) of PKHD1.@*Conclusion@#The three pedigrees were diagnosed as ARPKD caused by PKHD1 variants. Five types of variants were detected, c. 5935G>A and c. 11314C>T were the known pathogenic variants, while c. 5512T>C, c. 5428G>T and c. 3860T>G were not reported previously. Considering the complexity of the genetics and phenotypes of the cystic renal diseases, genetic diagnosis is crucial to give accurate etiological diagnosis, which may benefit the clinic management.