Impact of Combined Phototherapy and Melanocyte Transplantation on Indicators of Vitiligo Activity.

OBJECTIVE: To investigate the effect of phototherapy combined with melanocyte transplantation on the activity index of vitiligo. METHODS: One hundred twenty patients with stable vitiligo were selected and divided into 2 groups: phototherapy group (n = 60) and phototherapy combined with melanocyte therapy group (n = 60). Patients' vitiligo activity scores before and 6 months after treatment, patients' skin pigmentation responses 6 months after treatment, and patients' new Koebner cases 6 months after treatment were compared. The expression of tyrosinase and Melan-A in the skin samples was analyzed by immunohistochemistry. RESULTS: The effect of skin surface repigmentation in the observation group was better than that in the control group (p < .05). The expression of tyrosinase and Melan-A in the observation group was higher than that in the control group (p < .05), indicating that the combined treatment could enhance the function of melanocytes. After 6 months of treatment, the incidence of the Koebner phenomenon in the observation group was lower than that in the control group (p < .05). CONCLUSION: The combination of phototherapy and melanocyte transplantation can obviously improve the activity index of vitiligo, slow down the spread of white spots, reduce the formation of new white spots, and reduce the occurrence of the Koebner phenomenon.

Integrative analysis of proteomics and lipidomic profiles reveal the fat deposition and meat quality in Duroc × Guangdong small spotted pig.

Introduction: This study aims to explore the important factors affecting the characteristics of different parts of pork. Methods: Lipidomics and proteomics methods were used to analyze DAL (differential lipids) and DAPs (differential proteins) in five different parts (longissimus dorsi, belly meat, loin, forelegs and buttocks) of Duhua pig (Duroc × Guangdong small spotted pig), to identify potential pathways affecting meat quality, investigating fat deposition in pork and its lipid-protein interactions. Results: The results show that TG (triglyceride) is the lipid subclass with the highest proportion in muscle, and the pathway with the most significantly enriched lipids is GP. DAP clustered on several GO terms closely related to lipid metabolism and lipogenesis (lipid binding, lipid metabolism, lipid transport, and lipid regulation). In KEGG analysis, there are two main DAP aggregation pathways related to lipid metabolism, namely Fatty acid degradation and oxidative phosphorylation. In PPI analysis, we screened out 31 core proteins, among which NDUFA6, NDUFA9 and ACO2 are the most critical. Discussion: PC (phosphatidylcholine) is regulated by SNX5, THBS1, ANXA7, TPP1, CAVIN2, and VDAC2 in the phospholipid binding pathway. TG is regulated by AUH/HADH/ACADM/ACADL/HADHA in the lipid oxidation and lipid modification pathways. Potential biomarkers are rich in SFA, MUFA and PUFA respectively, the amounts of SFA, MUFA and PUFA in the lipid measurement results are consistent with the up- and down-regulation of potential biomarker lipids. This study clarified the differences in protein and lipid compositions in different parts of Duhua pigs and provided data support for revealing the interactions between pork lipids and proteins. These findings provide contributions to the study of intramuscular fat deposition in pork from a genetic and nutritional perspective.

Effects of dietary adenosine and adenosine 5'-monophosphate supplementation on carcass characteristics, meat quality, and lipid metabolism in adipose tissues of finishing pigs.

This study investigated the effects of adenosine (ADO) and adenosine 5'-monophosphate (AMP) supplementation on the growth performance, carcass characteristics, meat quality, and lipid metabolism in adipose tissues of finishing pigs. The pigs were allocated to three treatment groups: the control diet, 0.2%ADO diet, or 0.2%AMP diet. Compared with CON group (CON), both ADO and AMP groups increased in carcass straight length (P < 0.05) and decreased in drip loss (P < 0.05), while AMP group tended to increase in redness value (P = 0.05) and decreased in free amino acid content in longissimus thoracis (LT) muscle (P < 0.05). Additionally, ADO or AMP supplementation increased the ADO or AMP content in serum, adipose tissue, and LT muscle (P < 0.05), as well as the protein level of adenosine 2A receptor (A2a) in adipose tissue (P < 0.05). Moreover, both ADO and AMP groups showed an increase in the expression of lipolysis genes (ATGL and HSL) in adipose tissue (P < 0.05). Overall, AMP supplementation could improve meat quality, and ADO and AMP supplementation regulate the lipid metabolism of finishing pigs.

Effect of precision air supply cooling system with different cooling air speed on reproductive performance, stress status, immunoglobulin and fecal microbiota of lactating sows.

This study investigated the precision air supply cooling system with different cooling air speed on reproductive performance, stress status, immunity and intestinal microbiota of sows. A total of 36 sows were randomly assigned into three groups: control group (CON, n = 12) without cooling system, low cooling air speed (2.24 ± 0.11 m/s) group (LCAS, n = 12) with precision air supply cooling system, high cooling air speed (3.33 ± 0.11 m/s) group (HCAS, n = 12) with precision air supply cooling system. The average daily ambient temperature and relative humidity in experimental farrowing rooms were 28 °C-32 °C and 65%-90%, respectively, during daytime (8:00-20:00). The reproductive performance, respiratory rate (RR), body surface temperature (BST), rectal temperature (RT) salivary parameters and fecal microbiota of lactating sows were measured. The results showed that compared with CON group, average daily feed intake of sows and average daily gain of piglets were increased (P < 0.05) in LCAS and HCAS groups. The RR, BST and RT were decreased (P < 0.05) in LCAS and HCAS groups in wk 1 before parturition and wk 2 after parturition. The salivary cortisol and alpha amylase level of LCAS and HCAS sows were decreased (P < 0.05) on d 14 after parturition, while the salivary immunoglobulin A and immunoglobulin G level of LCAS and HCAS sows were increased (P < 0.05) on d 7 after parturition. The LCAS and HCAS sows showed higher relative abundance of Clostridium_sensu_stricto_1 in feces, which was positively associated with immunoglobulin A (P < 0.01). In conclusion, both low and high cooling air speed improved reproductive performance, alleviated stress status, and enhanced salivary immunoglobulins and beneficial fecal microbiota community in lactating sows, which indicated that precision air supply cooling system could be an effective cooling strategy for lactating sows in high ambient temperature.

NEDD4 E3 ubiquitin protein ligase serves an important role in cutaneous melanoma occurrence and development.

The present study aimed to discuss the effects and relative mechanisms of NEDD4 E3 ubiquitin protein ligase (NEDD4) in cutaneous melanoma (CMM) occurrence and development. Clinical cancer and adjacent normal tissues samples were collected to analyze pathological changes and protein expression of NEDD4. Moreover, small interfering (si)RNA was used to knockdown NEDD4 expression in SK-MEL-2 and Malme-3M cells. Cellular proliferation, apoptosis, invasiveness and migration were examined using colony formation, flow cytometric, Transwell and wound-healing assays, respectively. In addition, the relative mRNA and protein expression levels of NEDD4, notch receptor 1 (Notch1) and PTEN were evaluated via reverse transcription-quantitative (RT-q) PCR and western blotting. It was found that NEDD4 mRNA and protein expression were significantly upregulated (both P<0.01). Following NEDD4-knockdown, colony number was significantly decreased, while the apoptotic rate was significantly increased, the invasive cell number was significantly inhibited and the wound-healing capacity was significantly decreased. Following si-NEDD4 transfection, RT-qPCR and western blotting revealed that NEDD4 and Notch1 mRNA and protein expression levels were significantly downregulated, while those of PTEN were significantly upregulated in the SK-MEL-2 and Malme-3M cell lines. Collectively, the current results suggest that NEDD4-knockdown effectively suppressed CMM biological activity by regulating the Notch1/PTEN pathway in vitro.

Combining comparative genomic analysis with machine learning reveals some promising diagnostic markers to identify five common pathogenic non-tuberculous mycobacteria.

Non-tuberculous mycobacteria (NTM) can cause various respiratory diseases and even death in severe cases, and its incidence has increased rapidly worldwide. To date, it's difficult to use routine diagnostic methods and strain identification to precisely diagnose various types of NTM infections. We combined systematic comparative genomics with machine learning to select new diagnostic markers for precisely identifying five common pathogenic NTMs (Mycobacterium kansasii, Mycobacterium avium, Mycobacterium intracellular, Mycobacterium chelonae, Mycobacterium abscessus). A panel including six genes and two SNPs (nikA, benM, codA, pfkA2, mpr, yjcH, rrl C2638T, rrl A1173G) was selected to simultaneously identify the five NTMs with high accuracy (> 90%). Notably, the panel only containing the six genes also showed a good classification effect (accuracy > 90%). Additionally, the two panels could precisely differentiate the five NTMs from M. tuberculosis (accuracy > 99%). We also revealed some new marker genes/SNPs/combinations to accurately discriminate any one of the five NTMs separately, which provided the possibility to diagnose one certain NTM infection precisely. Our research not only reveals novel promising diagnostic markers to promote the development of precision diagnosis in NTM infectious, but also provides an insight into precisely identifying various genetically close pathogens through comparative genomics and machine learning.

A maternal high-fat/low-fiber diet impairs glucose tolerance and induces the formation of glycolytic muscle fibers in neonatal offspring.

PURPOSE: In our previous study, the maternal high-fat/low-fiber (HF-LF) diet was suggested to induce metabolic disorders and placental dysfunction of the dam, but the effects of this diet on glucose metabolism of neonatal offspring remain largely unknown. Here, a neonatal pig model was used to evaluate the effects of maternal HF-LF diet during pregnancy on glucose tolerance, transition of skeletal muscle fiber types, and mitochondrial function in offspring. METHODS: A total of 66 pregnant gilts (Guangdong Small-ear Spotted pig) at day 60 of gestation were randomly divided into two groups: control group (CON group; 2.86% crude fat, 9.37% crude fiber), and high-fat/low-fiber diet group (HF-LF group; 5.99% crude fat, 4.13% crude fiber). RESULTS: The maternal HF-LF diet was shown to impair the glucose tolerance of neonatal offspring, downregulate the protein level of slow-twitch fiber myosin heavy chain I (MyHC I), and upregulate the protein levels of fast-twitch fiber myosin heavy chain IIb (MyHC IIb) and IIx (MyHC IIx) in soleus muscle. Additionally, compared with the CON group, the HF-LF offspring showed inhibition of insulin signaling pathway and decrease in mitochondrial function in liver and soleus muscle. CONCLUSION: Maternal HF-LF diet during pregnancy impairs glucose tolerance, induces the formation of glycolytic muscle fibers, and decreases the hepatic and muscular mitochondrial function in neonatal piglets.

Placentae for Low Birth Weight Piglets Are Vulnerable to Oxidative Stress, Mitochondrial Dysfunction, and Impaired Angiogenesis.

Intrauterine growth restriction (IUGR) is associated with fetal mortality and morbidity. One of the most common causes of IUGR is placental insufficiency, including placental vascular defects, and mitochondrial dysfunction. In addition, a high level of oxidative stress induces placental vascular lesions. Here, we evaluated the oxidative stress status, mitochondrial function, angiogenesis, and nutrient transporters in placentae of piglets with different birth weights: <500 g (L), 500-600 g (LM), 600-700 g (M), and >700 g (H). Results showed that placentae from the L group had higher oxidative damage, lower adenosine triphosphate and citrate synthase levels, and lower vascular density, compared to those from the other groups. Protein expression of angiogenic markers, including vascular endothelial cadherin, vascular endothelial growth factor A, and platelet endothelial cell adhesion molecule-1, was the lowest in the L group placentae compared to the other groups. In addition, the protein levels of glucose transporters GLUT1 and GLUT3 were downregulated in the L group, compared to the other groups. Furthermore, oxidative stress induced by H2O2 inhibited tube formation and migration in porcine vascular endothelial cells. Collectively, placentae for lower birth weight neonates are vulnerable to oxidative damage, mitochondrial dysfunction, and impaired angiogenesis.

Effects of different maternal feeding strategies from day 1 to day 85 of gestation on glucose tolerance and muscle development in both low and normal birth weight piglets.

BACKGROUND: Maternal nutrition during gestation plays a vital role in fetal development. The effects of different maternal feeding strategies from day 1 to day 85 of gestation on glucose tolerance and muscle development in low and normal birth weight offspring were investigated by using 80 gilts randomly allotted to T1 and T2 groups and treated respectively with a gradual-increase (T1) and a convex transition (T2) feeding strategy, with no difference in total feed intake. RESULTS: T2 group was seen to have a higher percentage of piglets with birth weight less than 500 g, while T1 group was shown to have a higher percentage of piglets with birth weight over 700 g. Meanwhile, for both low and normal birth weight piglets, T1 group was higher than T2 group in terms of muscle free amino acid concentration, mRNA expression levels of muscle growth-related factors, relative muscle fiber number and cross-sectional area. We must emphasize that the T2 group was shown to improve glucose tolerance, slow-twitch muscle fiber protein levels, and muscle mitochondrial function only in low birth weight piglets. CONCLUSION: The convex transition feeding strategy can decrease the percentage of piglets with birth weight over 700 g, while improving glucose tolerance, slow-twitch muscle fiber protein levels, and muscle mitochondrial function in low birth weight piglets. Our findings provide new evidence for the potential importance of nutritional strategies during gestation, especially for improving the glucose tolerance and muscle development of low birth weight neonatal. © 2020 Society of Chemical Industry.

Precise editing of myostatin signal peptide by CRISPR/Cas9 increases the muscle mass of Liang Guang Small Spotted pigs.

Myostatin (MSTN), a member of the transforming growth factor-ß superfamily, is a negative regulator of muscle growth and development. Disruption of the MSTN gene in various mammalian species markedly promotes muscle growth. Previous studies have mainly focused on the disruption of the MSTN peptide coding region in pigs but not on the modification of the signal peptide region. In this study, the clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR associated protein 9 (Cas9) system was used to successfully introduce two mutations (PVD20H and GP19del) in the MSTN signal peptide region of the indigenous Chinese pig breed, Liang Guang Small Spotted pig. Both mutations in signal peptide increased the muscle mass without inhibiting the production of mature MSTN peptide in the cells. Histological analysis revealed that the enhanced muscle mass in MSTN+/PVD20H pig was mainly due to an increase in the number of muscle fibers. The expression of MSTN in the longissimus dorsi muscle of MSTN+/PVD20H and MSTNKO/PVD20H pigs was significantly downregulated, whereas that of myogenic regulatory factors, including MyoD, Myogenin, and Myf-5, was significantly upregulated when compared to those in the longissimus dorsi muscle of wild-type pigs. Meanwhile, the mutations also activated the PI3K/Akt pathway. The results of this study indicated that precise editing of the MSTN signal peptide can enhance porcine muscle development without markedly affecting the expression of mature MSTN peptide, which could exert other beneficial biological functions in the edited pigs.

Maternal Diet-Induced Obesity Compromises Oxidative Stress Status and Angiogenesis in the Porcine Placenta by Upregulating Nox2 Expression.

Maternal obesity is associated with placental oxidative stress. However, the mechanism underlying this association remains poorly understood. In the present study, a gilt obesity model was developed by exposure to different energy diets and used to investigate the role of NADPH oxidase 2 (Nox2) in the placenta. Specifically, 99 gilts (Guangdong Small-ear Spotted pig) at day 60 of gestation were randomly assigned to one of the following three treatments: low-energy group (L, DE = 11.50 MJ/kg), medium-energy group (M, DE = 12.41 MJ/kg), and high-energy group (H, DE = 13.42 MJ/kg), with 11 replicate pens per treatment and 3 gilts per pen. At the start of the study, maternal body weight and backfat thickness were not significantly different in the three treatments. After the study, data indicated that the H group had higher body weight and backfat thickness gain for gilts during gestation and lower piglet birth weight compared with the other two groups. Additionally, the H group showed glucolipid metabolic disorders and increased triglyceride and nonesterified fatty acid contents in the placenta of gilts. Compared with the L group, the H group exhibited lower mitochondrial biogenesis and increased oxidative damage in the placenta. Importantly, increased mRNA expression and protein abundance of Nox2 were observed for the first time in H group placentae. Furthermore, compared with the L group, the H group showed a decrease in the density of placental vessels and the protein levels of vascular endothelial cadherin (VE-cadherin), vascular endothelial growth factor A (VEGF-A), and phosphorylation of vascular endothelial growth factor receptor 2 (p-VEGFR2) as well as the immunostaining intensity of platelet endothelial cell adhesion molecule-1 (CD31). Our findings suggest that maternal high-energy diet-induced obesity increases placental oxidative stress and decreases placental angiogenesis possibly through the upregulation of Nox2.

Highly Efficient Generation of Pigs Harboring a Partial Deletion of the CD163 SRCR5 Domain, Which Are Fully Resistant to Porcine Reproductive and Respiratory Syndrome Virus 2 Infection.

Porcine reproductive and respiratory syndrome virus (PRRSV) 1 and 2 differ in their recognition of CD163. Substitution of porcine CD163 SRCR5 domain with a human CD163-like SRCR8 confers resistance to PRRSV 1 but not PRRSV 2. The deletion of CD163 SRCR5 has been shown to confer resistance to PRRSV 1 in vivo and both PRRSV 1 and 2 in vitro. However, the anti-PRRSV 2 activity of modifying the CD163 SRCR5 domain has not yet been reported. Here, we describe the highly efficient generation of two pig breeds (Liang Guang Small Spotted and Large White pigs) lacking a short region of CD163 SRCR5, including the ligand-binding pocket. We generated a large number of gene-edited Large White pigs of the F0 generation for use in viral challenge studies. The results of this study show that these pigs are completely resistant to infection by species 2 PRRSV, JXA1, and MY strains. There were no clinical symptoms, pathological abnormalities, viremia, or anti-PRRSV antibodies in the CD163 SRCR5-edited pigs compared to wild-type controls after viral challenge. Porcine alveolar macrophages (PAMs) isolated from CD163 SRCR5-edited Large White pigs also displayed resistance to PRRSV in vitro. In addition, CD163 SRCR5-edited PAMs still exhibited a cytokine response to PRRSV infection, and no significant difference was observed in cytokine expression compared to wild-type PAMs. Taken together, these data suggest that CD163 SRCR5-edited pigs are resistant to PRRSV 2, providing a basis for the establishment of PRRSV-resistant pig lines for commercial application and further investigation of the essential region of SRCR5 involved in virus infection.

Inactivation of Microcystis aeruginosa by DC glow discharge plasma: Impacts on cell integrity, pigment contents and microcystins degradation.

We proposed a method to inactivate M. aeruginosa by using discharge plasma taking at the gas-solution interface supplied by DC power. Multiple analysis techniques including fluorescence excitation-emission matrix (EEM) and flow cytometry (FCM) were used to reveal the inactivation mechanism of M. aeruginosa. The photosynthetic pigment contents including phycocyanin, chlorophyll and metabolites were examined quantitatively. The DC glow discharge plasma caused an increased level of reactive oxygen species (ROS), and the damage of M. aeruginosa cells are mainly attributed to the oxidative stress including OH attack and H2O2 oxidation. Our findings demonstrate that plasma oxidation is a promising technology for inactivation of M. aeruginosa cells with simultaneous removal of microcystins and so it may lead us to a new route to efficient treatment of cyanobacterial blooms.

Degradation of microcystin-LR in water by glow discharge plasma oxidation at the gas-solution interface and its safety evaluation.

Microcystin-LR (MC-LR) is one of the most commonly found microcystins (MCs) in fresh water and it poses danger to human health due to its potential hepatotoxicity. In the present study, we employed a novel method by using discharge plasma taking place at the gas-solution interface in gas atmosphere to degrade MC-LR in aqueous solution. The initial degradation rate of MC-LR was fastest under acidic conditions (5.41 ± 0.17 × 10(-3) mM min(-1) at pH 3.04) and decreased to 2.22 ± 0.11 × 10(-3) mM min(-1) and 0.912 ± 0.02 × 10(-3) mM min(-1) at pH 4.99 and 7.02, respectively. The effects of total soluble nitrogen (TN), total soluble phosphorus (TP) and natural organic matter (NOM) on the degradation efficiency were studied. The degradation rate was remarkably affected by TP and TN. Mass spectrometry was applied to identify the products of the reactions. Major degradation pathways are proposed according to the results of liquid chromatography/mass spectrometry (LC/MS) results. It suggests that the degradation of MC-LR is initiated via the attack of hydroxyl radicals on the conjugated carbon double bonds of Adda and on the benzene ring of Adda. Finally, the toxicity of intermediates or end-products from MC-LR degraded by this method was assessed using Caenorhabditis elegans. Our findings demonstrates that discharge plasma oxidation is a promising technology for degradation and removal of MC-LR and it may lead us to a new route to efficient treatment of other cyanotoxins from aqueous solutions.

Just enough measurement: a proposed paradigm for designing medical instrumentation.

Our research group hypothesizes that one way to provide low-cost healthcare delivery efficiently is through the use of a large number of inexpensive sensors that can provide meaningful medical data. Typical development of medical instrumentation pursues increased resolution and higher accuracy - accompanied by a corresponding increase in cost; it is no secret that high costs impose a heavy burden on healthcare. We seek to invert the adage that quality is more important than quantity by extracting high quality biomedical information from a large quantity of low-cost measurements, and to demonstrate this using measurement instrumentation developed in our lab for extra-clinical assessment and rehabilitation tools. This will be discussed in terms of our initial experiments in evaluating balance and postural stability. This is an area of critical clinical importance: 2.6 million non-fatal fall injuries in persons over age 65 resulted in direct health care costs of $19 billion (in 2000) in the U.S., and the number of persons over age 65 in the U.S. is projected to more than double between 2000 and 2030.