Effects of early intermittent maternal separation on behavior, physiological, and growth performance in piglets.

In pig production, the management of piglets by batch lactation due to the increase in litter sizes of sows may result in intermittent early neonatal maternal separation (NMS). We speculated that NMS may affect the piglets cognitive growth performance and health. To determine the extent of the effect, 12 litters of crossbred piglets (Large White × Duroc × Min-pig) were used in this trial. Piglets in the control (Con) group (n = 6) were given a standard feeding method during lactation. Piglets in the experimental group (n = 6) were subjected to the NMS model, in which sows were led out of the enclosure with food every day (8:00-11:00 and 13:00-16:00) starting from postnatal day (PND) 7. During the separation, the piglets were supplemented with milk. All experimental piglets were weaned on PND 35. The piglets were observed for aggression, play, mutual sniffing, and exploratory behavior on PNDs 7, 8, 21, 22, 34, 35, 38, 39, 51, 52, 64, and 65. Physiological indicators, namely serum adrenaline, cortisol, interleukin (IL)-1ß, IL-4, IL-6, and tumor necrosis factor (TNF)-α were measured on PNDs 35, 38, and 65, while piglet growth performance was evaluated during suckling and 1 month after weaning. The results showed that aggressive behavior in the MS group was significantly higher than that in the Con group (P < 0.05). Playful and mutual sniffing behaviors in the MS group were significantly lower than those in the Con group except for PNDs 38 and 39 (P < 0.05). Active exploratory behavior in the MS group was significantly higher than that in the Con group on PNDs 7 and 8, and PNDs 21 and 22 (P < 0.05). The frequency of belly-nosing behavior was significantly higher in the MS group than that in the Con group except for PNDs 64 and 65 (P < 0.05). Compared with the Con group, epinephrine, IL-1ß, IL-6, and TNF-α concentrations on PNDs 35, 38, and 65 were significantly increased in the MS group (P < 0.01), while IL-4 concentration was significantly decreased (PND 35: P < 0.05; PNDs 38 and 65: P < 0.01). Compared with the Con group, the piglet diarrhea rate in the MS group during suckling was significantly increased (P < 0.01), the weaning weight was significantly decreased (P < 0.05), and it had no significant effect on the body weight at the end of the trial (P > 0.05). In conclusion, the early intermittent NMS created stress and affected the growth performance of piglets during suckling. However, the growth rate was improved by compensatory measures during late weaning.

Although management methods, such as split-suckling and foster care, in pig production can improve piglet survival rates, these methods inevitably lead to neonatal maternal separation which is an early stress on the body, and can have serious negative effects on the body. In this experiment, we investigated the effect level of neonatal maternal separation on behavior, physiology, and growth performance of piglets. The study found that early intermittent maternal separation leads to anxiety and behavioral changes in piglets, negatively affecting diarrhea rates and weaning weights in suckling piglets, but the effects on growth performance in lactating piglets can be ameliorated during the nursing period.

Identification of a novel tumour microenvironment-based prognostic biomarker in skin cutaneous melanoma.

Skin cutaneous melanoma (SKCM) is one of the most destructive skin malignancies and has attracted worldwide attention. However, there is a lack of prognostic biomarkers, especially tumour microenvironment (TME)-based prognostic biomarkers. Therefore, there is an urgent need to investigate the TME in SKCM, as well as to identify efficient biomarkers for the diagnosis and treatment of SKCM patients. A comprehensive analysis was performed using SKCM samples from The Cancer Genome Atlas and normal samples from Genotype-Tissue Expression. TME scores were calculated using the ESTIMATE algorithm, and differential TME scores and differentially expressed prognostic genes were successively identified. We further identified more reliable prognostic genes via least absolute shrinkage and selection operator regression analysis and constructed a prognostic prediction model to predict overall survival. Receiver operating characteristic analysis was used to evaluate the diagnostic efficacy, and Cox regression analysis was applied to explore the relationship with clinicopathological characteristics. Finally, we identified a novel prognostic biomarker and conducted a functional enrichment analysis. After considering ESTIMATEScore and tumour purity as differential TME scores, we identified 34 differentially expressed prognostic genes. Using least absolute shrinkage and selection operator regression, we identified seven potential prognostic biomarkers (SLC13A5, RBM24, IGHV3OR16-15, PRSS35, SLC7A10, IGHV1-69D and IGHV2-26). Combined with receiver operating characteristic and regression analyses, we determined PRSS35 as a novel TME-based prognostic biomarker in SKCM, and functional analysis enriched immune-related cells, functions and signalling pathways. Our study indicated that PRSS35 could act as a potential prognostic biomarker in SKCM by investigating the TME, so as to provide new ideas and insights for the clinical diagnosis and treatment of SKCM.

SHARPIN regulates cell proliferation of cutaneous basal cell carcinoma via inactivation of the transcriptional factors GLI2 and c­JUN.

SHANK­associated RH domain­interacting protein (SHARPIN) is a component of the linear ubiquitin chain assembly complex that can enhance the NF­κB and JNK signaling pathways, acting as a tumor­associated protein in a variety of cancer types. The present study investigated the role of SHARPIN in cutaneous basal cell carcinoma (BCC). Human BCC (n=26) and normal skin (n=5) tissues, and BCC (TE354.T) and normal skin (HaCaT) cell lines were used to evaluate SHARPIN expression level using immunohistochemistry and western blotting, respectively. A lentivirus carrying SHARPIN­targeting or negative control short hairpin RNA was infected into TE354.T cells, and the infected stable cells were assayed to analyze tumor cell proliferation, cell cycle, apoptosis, migration and invasion by Cell Counting Kit­8 and 5­ethynyl­2'­deoxyuridine incorporation assays, flow cytometry and Transwell assays. Western blotting was performed to assess the protein expression levels of gene signaling in SHARPIN­silenced BCC cells. SHARPIN protein expression levels were downregulated or absent in BCC cancer nests and precancerous lesions compared with normal skin samples. In addition, SHARPIN expression levels were lower in TE354.T cells compared with HaCaT cells. SHARPIN shRNA enhanced tumor cell proliferation and the S phase of the cell cycle, whereas BCC cell apoptotic rates, and migratory and invasive abilities were not significantly altered. The expression levels of cyclin D1, cyclin­dependent kinase 4, phosphorylated­c­JUN and GLI family zinc finger 2 proteins were increased, whereas Patched 1 (PTCH1) and PTCH2 were decreased in the SHARPIN­shRNA­infected BCC cells. Therefore, the present results suggested that SHARPIN may act as a tumor suppressor during BCC development.

A Strategy for Quality Control of Vespa magnifica (Smith) Venom Based on HPLC Fingerprint Analysis and Multi-Component Separation Combined with Quantitative Analysis.

As a folk medicine of the Jingpo minority in Yunnan province, the venom of Vespa magnifica has been commonly used for the treatment of rheumatoid arthritis. Quality standardization of the wasp venom is a necessary step for its pharmaceutical research and development. To control the quality of the wasp venom, a method based on high-performance liquid chromatography (HPLC) was developed for chemical fingerprint analysis. In the chromatographic fingerprinting, chemometrics procedures, including similarity analysis (SA), hierarchical clustering analysis (HCA), and principal component analysis (PCA), were applied to classify 134 batches (S1-S134) of wasp venom from different origins. The HPLC fingerprint method displayed good precision (Relative standard deviation, RSD < 0.27%), stability (in 16 h, RSD < 0.34%), and repeatability (RSD < 1.00%). Simultaneously, four compounds (VMS1, VMS2, VMS3, and VMS4) in the wasp venom were purified and identified. VMS1 was 5-hydroxytryptamine, and the other compounds were three peptides that were sequenced as follows: Gly-Arg-Pro-Hyp-Gly-Phe-Ser-Pro-Phe-Arg-Ile-Asp-NH2 (VMS2), Ile-Asn-Leu-Lys-Ala-Ile-Ala-Ala-Leu-Ala-Lys-Lys-Leu-Leu-NH2 (VMS3), and Phe-Leu-Pro-Ile-Ile-Gly-Lys-Leu-Leu-Ser-Gly-Leu-Leu-NH2 (VMS4). The quantifications for these components were 110.2 mg/g, 26.9 mg/g, 216.3 mg/g, and 58.0 mg/g, respectively. The results of this work indicated that the combination of the chemical fingerprint and quantitative analysis offers a reasonable way to evaluate the quality of wasp venom.

[Effect of 0.9-ms 1064-nm Nd:YAG laser combined with itraconazole for treatment of toenail onychomycosis].

OBJECTIVE: To evaluate the efficacy and safety of 0.9-ms 1064-nm Nd:YAG laser alone or combined with itraconazole for treatment of toenail onychomycosis. METHODS: A total of 37 patients with onychomycosis (178 toenails) were randomly assigned to groups A and B, and each group was further divided into different subgroups according to the Scoring Clinical Index of Onychomycosis (SCIO) and Onychomycosis Severity Index (OSI) scoring. All the patients were treated with 0.9-ms Nd:YAG laser once a week for 8 times. The patients in group A were treated with laser alone, and those in group B were treated with laser combined with itraconazole. The clinical effect, clinical scores, appearance of the toenails and adverse reactions in the two groups were analyzed, and the patients' satisfaction rate was also investigated. RESULTS: At the 12th months of follow-up, the clinical response rate and mycological cure rate in group A were 31.33% and 30.00%, respectively, similar to the rates in group B (35.79% and 41.18%, respectively) (P>0.05). After the treatments, the SCIO and OSI scores showed no significant changes in group A (P>0.05) but both increased significantly in group B (P<0.05). The response rates did not differ significantly among the subgroups with SCIO<12 or with OSI<16 (P>0.05), but showed significant differences among the subgroups with SCIO≥12 or with OSI≥16 (P<0.05). Of the total of 178 toenails, 33.71%, 74.72% and 70.79% toenails showed improvements in terms of clear nail growth, shape and color, respectively. The overall patients' satisfaction rate was 62.16%, and no adverse reactions related with the therapy were recorded in these patients. CONCLUSION: For treatment of toenail onychomycosis, 0.9-ms 1064-nm Nd:YAG laser can effectively improve the aesthetic appearance of the toenails, and a combined treatment with Nd:YAG laser and itraconazole can be better option in severe cases of onychomycosis.

[Efficacy and safety of long pulse 1064 nm Nd:YAG laser for treatment of onychomycosis of the toenails].

OBJECTIVE: To evaluate the efficacy and safety of long pulse 1064 nm Nd:YAG laser therapy in the treatment of onychomycosis of the toenails. METHODS: A total of 104 patients with onychomycosis (461 toenails) were divided by age into ≥60 years group and <60 years group, and each group was further divided into subgroups according to Scoring Clinical Index of Onychomycosis (SCIO) scoring and the location of the compromised toenails. All the toenails were treated with 10 to12 sessions of long pulse 1064 nm Nd:YAG laser therapy at the interval of 1 week. All the patients were followed up for 48 weeks after the initial treatment to assess the clinical efficacy and adverse reactions. RESULTS: The overall clinical response rate in these patients was 72.5% by the end of the 48-week follow-up. In patients aged <60 years, the clinical response rate and mycological cure rate were significantly higher than the rates in patients aged ≥60 years (P<0.05). No significant differences were observed in the response rates between different SCIO subgroups (P>0.05); the 2nd to 4th toenails showed better outcomes after the therapy than the 1st and 5th toenails (P<0.05). No adverse reactions related with the therapy were recorded in these patients. CONCLUSION: Long pulse 1064 nm Nd:YAG laser is an effective and safe approach for treatment of onychomycosis of the toenails.

[Apoptosis effects of drug sensitivity leukemia cells induced by nano-realgar].

OBJECTIVE: To explore apoptosis-inducing effects of realgar nanoparticle (nano-realgar) on drug-sensitive leukemia cells. METHOD: Preparation of nano-realgar was mechanical milled using a high-energy planetary ball mill. Using drug-sensitive leukemia cells (K562) as target cells, MTT assay was used to detect the proliferating activity of K562 cells, and the cellular apoptosis was investigated with double staining of FITC-Annexin V and propidium iodide (PI) by flow cytometry. Flow cytometry (FCM) was employed to detect expression of intracellular Bax, Bcl-2, P-53 protein and the activity of Caspase-3. RESULT: The raw realgar was made to ultra-fine powder by ball milling, and the average diameter of the nanoparticle was (72.72 +/- 22.18) nm measured with electron microscopes. Nano-realgar significantly inhibited the proliferation of K562 cells, Treated for 24, 48 and 72 hours, the 50% inhibitory concentration (IC50) was 43.48, 20.52, 16.07 mg x L(-1). After exposure to 20 mg x L(-1) and 50 mg x L(-1) nano-realgar for 48 hours, the apoptosis of K562 cells detected by Annexin V/PI staining was increased, the apoptotic rate of K562 cells was 10. 52% and 73.25%. After the target cells were treated with 20 mg x L(-1) and 50 mg x L(-1) nano-realgar for 48 h, the expression of P-53, Bax, Bcl-2 markedly increased in a time and dose-dependent manner. After administration of 20 mg x L(-1) and 50 mg x L(-1) nano-realgar for 48 h, the percentage of BCRP+, P-gp+ and co-expressing P-gp and BCRP cell population in K562 cells incrased dramatically. CONCLUSION: Nano-Realgar significantly induced apoptosis of drug-sensitive leukemia cells.