Hypoxia Modulates Melanoma Cells Proliferation and Apoptosis via miRNA-210/ISCU/ROS Signaling.

In this study, luciferase reporter assay was used to establish the relationship between miR-210 and ISCU. This research was performed on both cell lines (A2058, G361, and 293T) and tissue samples. We found that miR-210 was upregulated in hypoxia and was elevated in melanoma in comparison with adjacent normal tissues. Expression of ISCU protein was decreased in melanoma tissues, and ISCU gene is a direct target of miR-210. ISCU knockdown with miR-210 enhanced ROS production. The results of our study showed that miR-210/ISCU/ROS axis can serve as a novel therapeutic target in melanoma.

Mussel-Inspired Hydrogels for Fluoride Delivery and Caries Prevention.

Fluoride agents hold promise for the repair and prevention of caries lesions, but their interaction with enamel is often hampered and diminished because of the dynamic wet environment in the oral cavity, which affects the efficacy of fluoride delivery and limits treatment success. We herein developed a mussel-inspired wet adhesive fluoride system (denoted TS@NaF) fabricated by the self-assembly of tannic acid (TA), silk fibroin (SF), and sodium fluoride (NaF). TS@NaF demonstrated remarkable biological stability and biocompatibility, showed reliable wet adhesion, released fluoride ions (F-) topically, and induced significant deposition of calcium fluoride (CaF2) onto enamel in vitro. Furthermore, TS@NaF provided an anticaries effect in vitro and induced a detectable increase in enamel mineral density. Advanced fluoride-releasing bioadhesives are therefore promising candidates for caries prevention and highlight the great potential of mussel-inspired dental materials in clinical applications.

[Prenatal genetic diagnosis of the fetuses with isolated corpus callosum abnormality].

Objective: To explore the application value of chromosome karyotype analysis, chromosomal microarray analysis (CMA) and whole exome sequencing (WES) in prenatal diagnosis of isolated corpus callosum abnormality (CCA) fetus. Methods: Fetuses diagnosed with isolated CCA by ultrasound and MRI and receiving invasive prenatal diagnosis in Guangzhou Women and Children's Medical Center and Qingyuan People's Hospital from January 2010 to April 2021 were selected. Karyotype analysis and/or CMA [or copy number variation sequencing (CNV-seq)] were performed on all fetal samples, and WES was performed on fetal samples and their parents whose karyotype analysis and/or CMA (or CNV-seq) results were not abnormal. Results: Among 65 fetuses with isolated CCA, 38 cases underwent karyotype analysis, and 3 cases were detected with abnormal karyotypes, with a detection rate of 8% (3/38). A total of 49 fetuses with isolated CCA underwent CMA (or CNV-seq) detection, and 6 cases of pathogenic CNV were detected, the detection rate was 12% (6/49). Among them, the karyotype analysis results were abnormal, and the detection rate of further CMA detection was 1/1. The karyotype results were normal, and the detection rate of further CMA (or CNV-seq) detection was 14% (3/21). The detection rate of CMA as the first-line detection technique was 7% (2/27). A total of 25 fetuses with isolated CCA with negative results of karyotyping and/or CMA were tested by WES, and 9 cases (36%, 9/25) were detected with pathogenic genes. The gradient genetic diagnosis of chromosomal karyotyping, CMA and WES resulted in a definite genetic diagnosis of 26% (17/65) of isolated CCA fetuses. Conclusions: Prenatal genetic diagnosis of isolated CCA fetuses is of great clinical significance. The detection rate of CMA is higher than that of traditional karyotyping. CMA detection could be used as a first-line detection technique for fetuses with isolated CCA. WES could increase the pathogenicity detection rate of fetuses with isolated CCA when karyotype analysis and/or CMA test results are negative.

Effect of integrative medicines on 28-day mortality from sepsis: a systematic review and network meta-analysis.

OBJECTIVE: The aim is to perform a network meta-analysis to evaluate the effect of different Chinese medicines combined with Western medicine treatment (WMT) on the 28-day mortality of patients with sepsis. MATERIALS AND METHODS: We searched multiple databases for randomized controlled trials (RCTs), using keywords such as sepsis, Shenfu, Shenmai, Shengmai, Dahuang Fuzi, Xuebijing, and mortality. The databases we searched included the China Knowledge Network (CNKI), Wanfang database, the Chinese scientific journal database (VIP), the Chinese biomedical literature database (CBM), PubMed, Embase and Cochrane. The collection time was from inception through September 22, 2020. Two researchers independently screened the articles, extracted data, and assessed the quality of the evidence. Stata15 was used for network meta-analysis, and Excel was used to summarize the list of adverse reactions. RESULTS: Twenty-nine studies with a total of 3201 patients were included in this study. Combined with WMT, Shenfu injection [OR = 0.63, 95% CI = (0.47, 0.83)] and Xuebijing injection [OR = 0.71, 95% CI = (0.52, 0.96)] can significantly reduce 28-day mortality among patients with sepsis, while Shenmai injection [OR = 0.40, 95% CI = (0.16, 1.03)] and Dahuang Fuzi decoction [OR = 1.00, 95% CI = (0.48, 2.10)] cannot significantly improve on the 28-day mortality. The efficacy ranking is as follows: Shenmai group (level of evidence: low) > Shenfu group (level of evidence: low) > Xuebijing group (level of evidence: very low) > WMT group > Dahuangfuzi group (level of evidence: very low). CONCLUSIONS: Combined with WMT, Shenfu injection or Xuebijing injection can reduce the 28-day mortality among patients with sepsis. The Shenfu group had the best effect on outcomes, and its level of evidence was higher than that of the Xuebijing group.

[Analysis of families with fetal congenital abnormalities but negative prenatal diagnosis by whole exome sequencing].

Objective: To evaluate the value of whole exome sequencing (WES) in prenatal clinical application. Methods: A total of 1 152 cases of congenital abnormal [including structural malformation, nuchal translucency (NT) thickening and intrauterine growth restriction] with traditional prenatal diagnosis [including G-band karyotype analysis and chromosome microarray analysis (CMA)] negative were analyzed. The congenital abnormal fetuses were divided into retrospective group and prospective group according to the time of WES detection, that is whether the pregnancy termination or not. According to the specific location of fetal malformation and their family history, the cohort was divided into subgroups. The clinical prognosis of all fetuses were followed up, and the effect of WES test results on pregnancy decision-making and clinical intervention were analyzed. According to the follow-up results, the data of fetuses with new phenotypes in the third trimester or after birth were re-analyzed. Results: Among 1 152 families who received WES, 5 families were excluded because of nonbiological parents. Among the remaining 1 147 families, 152 fetuses obtained positive diagnosis (13.3%,152/1 147), including 74 fetuses in the retrospective group (16.1%,74/460) and 78 fetuses in the prospective group (11.4%,78/687). In fetuses with negative CMA and G-band karyotype analysis results but new phenotypes in the third trimester or after birth, the positive rate by WES data re-analysis was 4.9% (8/163). A total of 34 (21.3%, 34/160) fetuses were directly affected by the corresponding positive molecular diagnosis. Among 68 cases of live births with diagnostic variation grade 4, 29 cases (42.7%, 29/68) received appropriate medical intervention through rapid review of WES results. Conclusions: WES could increase the detection rate of abnormal fetuses with negative G-banding karyotype analysis and CMA by 13.3%. Prenatal WES could guide pregnancy decision-making and early clinical intervention. It might be an effective strategy to pay attention to the special follow-up of the third trimester and postnatal fetus and to re-analyze the WES data.

[Management strategy for the resumption of regular diagnosis and treatment in gastrointestinal surgery department during the outbreak of coronavirus disease 2019 (COVID-19)].

Acute abdomen, abdominal trauma, gastrointestinal bleeding and gastrointestinal tumors are the main conditions that are routinely treated in gastrointestinal surgery department with high incidence and critical condition. These conditions need emergency or selective operations. During the outbreak of the coronavirus disease 2019 (COVID-19), it's a great challenge for us to meet the patients' requirement under the situation. As the COVID-19 was brought under control in China, the Department of General Surgery in Nanfang Hospital resumed regular medical services gradually. Based on our clinical practice, the four major measures of strengthening pre-hospital screening, perioperative prevention and control, medical staff protection, and ward management were adopted. These main measures include the strict implementation of the appointment system and triage system before admission; the conduction of epidemiological and preliminary screening of viral nucleic acids; the chest CT examination during the perioperative period to re-screen COVID-19; the reduction of the risk of droplets and aerosol transmission; the minimally invasive surgery combined with enhanced recovery program in order to reduce patient's susceptibility and shorten the length of postoperative hospital stay; the reinforcement of specific infection control training for medical staff; the strict implementation of hierarchical protection; the establishment of gastrointestinal surgery prevention and control system; the rehearsal of emergency exercise; the installation of quarantine wards; the screening and management of family care-givers; the strict disinfection of environment and materials. Our preliminary practice shows that following the work guidelines issued by the Guangdong Province COVID-19 Prevention and Control Office and adopting precise management strategies in combination with the specific clinical features of gastrointestinal surgery, it is possible to safely resume regular care for the patients and comply to epidemic control at the same time.

AXIN2 gene silencing reduces apoptosis through regulating mitochondria-associated apoptosis signaling pathway and enhances proliferation of ESCs by modulating Wnt/ß-catenin signaling pathway.

OBJECTIVE: Embryonic stem cells (ESCs) mainly originate from totipotent cells in early-stage of mammalian embryo and could proliferate in a manner of un-limitation. This study aimed to investigate roles of Axin2 in proliferation of ESCs and explore the associated mechanisms. MATERIALS AND METHODS: Axis inhibition protein 2 (AXIN2) over-expression (LV5-AXIN2) and AXIN2 RNA interfere (LV3-AXIN2-RNAi) vectors were structured and transfected into H9 cells. 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT) was used to evaluate cell proliferative activity. Flow cytometry analysis was employed to measure apoptosis of H9 cells. AXIN2, ß-catenin, transcription factor 4 (TCF4), c-myc, c-jun and Cyclin D mRNA levels and protein expressions were determined using quantitative real-time PCR (qRT-PCR) and Western blotting assay. RESULTS: LV5-AXIN2 and LV3-AXIN2-RNAi were successfully structured with higher transfecting efficacy. AXIN2 gene silencing remarkably increased proliferative activity and AXIN2 treatment significantly induced apoptosis of H9 cells, comparing with blank vector group (p<0.05). AXIN2 gene silencing significantly enhanced B-cell lymphoma-2 (Bcl-2) expression and remarkably inhibited cleaved caspase-3 expression comparing to that in blank vector group (p<0.05). AXIN2-RNAi treatment significantly enhanced and AXIN2 over-expression significantly reduced ß-catenin and TCF4 expression, comparing to that in blank vector group (p<0.05). AXIN2 gene silence activated down-stream molecules of Wnt/ß-catenin signaling pathway, including c-jun, c-myc, and Cyclin D1 (p<0.05). CONCLUSIONS: AXIN2 gene silencing reduced apoptosis by regulating mitochondria-associated apoptosis signaling pathway and enhanced proliferation by modulating molecules in Wnt/ß-catenin signaling pathway. Therefore, targeting of aberrant apoptosis and AXIN2 might be a novel clinical strategy to inhibit aging and enhance self-renewal of ESCs.

Tumor invasion front morphology: a novel prognostic factor for intrahepatic cholangiocarcinoma.

OBJECTIVE: To explore the prognostic value of the morphology of tumor invasion front (TIF) in Intrahepatic Cholangiocarcinoma (ICC). PATIENTS AND METHODS: Seventy-four ICC patients with complete clinicopathological data and follow-up information were enrolled in our study. The most typical morphology of TIF of each case will be classified as low-grade group or high-grade group after evaluation by two pathologists. The clinicopathological characteristics, disease-free survival (DFS), and overall survival (OS) were compared between the two groups. RESULTS: A total of 26 (35.3%) patients were assigned to low-grade group, while 48 (64.7%) patients were assigned to high-grade group. High-grade group was associated with higher CA19-9 (p=0.032), poor differentiation (p=0.050), larger tumor diameter (p=0.016), advanced T staging (p=0.048), and higher incidence of lymph node (LN) metastasis (0.014). No significant associations were found in demographic and clinical characteristics between the two groups. On multivariable analysis, high-grade group was a significant independent predictor of worse DFS (HR=0.433, 95% CI=0.235-0.800, p=0.002) and OS (HR=0.363, 95% CI=0.187-0.704, p=0.003). CONCLUSIONS: High-grade morphology of TIF was an independent prognostic factor of ICC.

Gadolinium Deposition in Brain: Current Scientific Evidence and Future Perspectives.

In the past 4 years, many publications described a concentration-dependent deposition of gadolinium in the brain both in adults and children, seen as high signal intensities in the globus pallidus and dentate nucleus on unenhanced T1-weighted images. Postmortem human or animal studies have validated gadolinium deposition in these T1-hyperintensity areas, raising new concerns on the safety of gadolinium-based contrast agents (GBCAs). Residual gadolinium is deposited not only in brain, but also in extracranial tissues such as liver, skin, and bone. This review summarizes the current evidence on gadolinium deposition in the human and animal bodies, evaluates the effects of different types of GBCAs on the gadolinium deposition, introduces the possible entrance or clearance mechanism of the gadolinium and potential side effects that may be related to the gadolinium deposition on human or animals, and puts forward some suggestions for further research.

Prognostic factors for treatment success in patients with multidrug-resistant tuberculosis in China.

OBJECTIVE: To examine the clinical outcomes and associated prognostic factors among patients with multidrug-resistant tuberculosis (MDR-TB) in China. METHODS: This retrospective study involved 243 patients with MDR-TB. All patients received standard regimens containing para-amino salicylic acid (PAS) and/or cycloserine (CS). The demographic, social and clinical characteristics of patients were recorded and the patients were followed up for 24 months. RESULTS: Treatment success was closely associated with young age, non-farming occupations, shorter history or smoking, normal urine results, initial MDR-TB treatment regimen, increased haemoglobin, direct bilirubin, uric acid and thyroid stimulating hormone (TSH) levels, and lower white blood cell, neutrophil and blood platelet counts (all P < 0.05). On multivariable analysis, increased haemoglobin (hazard ratio [HR] 1.019, 95%CI 1.007-1.032, P = 0.002) and TSH levels (HR 1.002, 95%CI 1.006-1.039, P = 0.008), normal urine results (HR 1.541, 95%CI 1.008-2.358, P = 0.046) and initial MDR-TB treatment regimen (HR 2.238, 95%CI 1.090-4.597, P = 0.028) were prognostic factors for treatment success in MDR-TB. CONCLUSIONS: Higher haemoglobin and TSH levels, normal urine results and initial MDR-TB treatment regimen might predict successful treatment of MDR-TB.

Whole exome sequencing as a diagnostic adjunct to clinical testing in fetuses with structural abnormalities.

OBJECTIVES: To evaluate the diagnostic yield of prenatal whole exome sequencing (WES) for monogenic disorders in fetuses with structural malformations and normal results on cytogenetic testing, and to describe information on pathogenic variants that is provided by WES. METHODS: Karyotyping, chromosomal microarray analysis (CMA) and WES were performed sequentially on stored samples from a cohort of 3949 pregnancies with fetal structural abnormalities detected on ultrasound and/or magnetic resonance imaging, referred between January 2011 and December 2015. Diagnostic rates of the three techniques were investigated overall, for phenotypic subgroups and for proband-only vs fetus-mother-father samples. Information on pathogenic variants was identified by WES. RESULTS: Overall, 18.2% (720/3949) of fetuses had an abnormal karyotype. Pathogenic copy number variants were detected on CMA in 8.2% (138/1680) of fetuses that had a normal karyotype result. WES performed on a subgroup of 196 fetuses with normal CMA and karyotype results revealed the putative genetic variants responsible for the abnormal phenotypes in 47 cases (24%). The molecular diagnosis rates for fetus-mother-father and proband-only samples were 26.5% (13/49) and 23.1% (34/147), respectively. Variants of uncertain significance were detected in 12.8% (25/196) of fetuses, of which 22 were identified in the fetal proband-only group (15%; 22/147) and three in the fetus-mother-father group (6.1%; 3/49). The incidental finding rate was 6.1% (12/196). CONCLUSIONS: WES is a promising method for the identification of genetic variants that cause structural abnormalities in fetuses with normal results on karyotyping and CMA. This enhanced diagnostic yield has the potential to improve the clinical management of pregnancies and to inform better the reproductive decisions of affected families. Copyright © 2017 ISUOG. Published by John Wiley & Sons Ltd.

PET/MR Imaging: New Frontier in Alzheimer's Disease and Other Dementias.

Alzheimer's disease (AD) is the most common form of dementia; a progressive neurodegenerative disease that currently lacks an effective treatment option. Early and accurate diagnosis, in addition to quick elimination of differential diagnosis, allows us to provide timely treatments that delay the progression of AD. Imaging plays an important role for the early diagnosis of AD. The newly emerging PET/MR imaging strategies integrate the advantages of PET and MR to diagnose and monitor AD. This review introduces the development of PET/MR imaging systems, technical considerations of PET/MR imaging, special considerations of PET/MR in AD, and the system's potential clinical applications and future perspectives in AD.

Effect of feeding Rumen-protected capsule containing niacin, K2SO4, vitamin C, and gamma-aminobutyric acid on heat stress and performance of dairy cows.

This study was conducted to evaluate the effects of supplemental rumen-protected capsule (RPC) on animal performance, serological indicators, and serum heat shock protein 70 (HSP70) of lactating Holstein cows under heat stress (HS). During summer months, 30 healthy multiparous lactating Holstein cows with a parity number of 3.1 ± 0.44, 70 ± 15 d in milk, an average body weight of 622 ± 62kg, and an average milk yield of 32.28 ± 0.96kg/d, were used. The cows were randomly allocated to two groups: a control group and an RPC-supplemented group (0.13373kg K2SO4, 0.02488kg vitamin C, 0.021148kg niacin, and 0.044784kggamma-aminobutyric acid per cow). During the 42-d experiment, ambient air temperature and relative humidity inside and outside the barn were recorded hourly every day for the determination of temperature-humidity index (THI). Milk and blood samples were collected every week, and body weight and body condition scoring were measured on day 0. Based on the THI values, the animals had moderate HS. On day 42, the RPC group had lower HSP70, adrenocorticotropic hormone (P = 0.0001), lactate dehydrogenase (P = 0.0338), and IL-6 (P = 0.0724) levels than the control group, with no significant differences in creatine kinase, glucocorticoid, or IL-2 levels. Milk yield, energy-corrected milk, and dry matter intake were higher in RPC than in the control group (P = 0.0196). There were no significant differences in milk fat or daily protein levels between the two groups; however, daily protein and milk fat levels were higher in the RPC group than in the control group (P = 0.0114 and P = 0.0665, respectively). Somatic cell counts were no different between the two groups. In conclusion, RPC may alleviate HS and improve dairy cow performance.

[Primary culture of cat intestinal epithelial cell and construction of its cDNA library].

Objective To establish the primary cat intestinal epithelial cells (IECs) culture methods and construct the cDNA library for the following yeast two-hybrid experiment, so as to screen the virulence interaction factors among the final host. Methods The primary cat IECs were cultured by the tissue cultivation and combined digestion with collagenase XI and dispase I separately. Then the cat IECs cultured was identified with the morphological observation and cyto-keratin detection, by using goat anti-cyto-keratin monoclonal antibodies. The mRNA of cat IECs was isolated and used as the template to synthesize the first strand cDNA by SMART™ technology, and then the double-strand cDNAs were acquired by LD-PCR, which were subsequently cloned into the plasmid PGADT7-Rec to construct yeast two-hybrid cDNA library in the yeast strain Y187 by homologous recombination. Matchmaker™ Insert Check PCR was used to detect the size distribution of cDNA fragments after the capacity calculation of the cDNA library. Results The comparison of the two cultivation methods indicated that the combined digestion of collagenase XI and dispase I was more effective than the tissue cultivation. The cat IECs system of continuous culture was established and the cat IECs with high purity were harvested for constructing the yeast two-hybrid cDNA library. The library contained 1.1×106 independent clones. The titer was 2.8×109 cfu/ml. The size of inserted fragments was among 0.5-2.0 kb. Conclusion The yeast two-hybrid cDNA library of cat IECs meets the requirements of further screen research, and this study lays the foundation of screening the Toxoplasma gondii virulence interaction factors among the cDNA libraries of its final hosts.