miR-21 promotes proliferation and inhibits apoptosis of hepatic stellate cells through targeting PTEN/PI3K/AKT pathway.

To investigate the effect of microRNA 21 (miR-21) on hepatic stellate cells (HSCs) proliferation and apoptosis, and further to study its potential mechanisms. LX-2 cells were divided into miR-21 mimic group (Mimic), miR-21 mimic negative control group (NM), miR-21 inhibitor group (Inhibitor), miR-21 inhibitor negative control group (NC), and blank control group (Control). The cell proliferation was detected by CCK-8 assay and the cell migration and invasion were detected by scratch and transwell assay. Cell cycle and apoptosis were detected by flow cytometry. The levels of interleukin (IL)-6, tumor necrosis factor (TNF)-α, and transforming growth factor (TGF)-ß1 were detected by enzyme-linked immunosorbent assay (ELISA). Proliferation, apoptosis, and phosphatase and tensin homolog (PTEN)/phosphoinositide 3-kinase (PI3K)/protein kinase B (AKT) signaling pathway related genes and proteins were detected by quantitative real-time polymerase chain reaction (qRT-PCR) and western blot, respectively. The cells proliferation, migration, and invasion were promoted in Mimic group. The levels of IL-6, TNF-α, and TGF-ß1 were increased after miR-21 administration. The expression of α-smooth muscle actin (SMA) and collagen 1 (Colla1) were increased, while Bax/B-cell lymphoma (Bcl)-2 ratio and programed cell death 4 (PDCD4) were reduced after miR­21 treatment. Meanwhile, the mRNA and protein expression of PTEN were reduced and PI3K/AKT pathway been promoted. Our study demonstrated that miR-21 could promote proliferation and inhibit apoptosis of HSCs, and its mechanism may be related to PTEN/PI3K/AKT pathway.

Prolactin levels as a criterion to differentiate between psychogenic non-epileptic seizures and epileptic seizures: A systematic review.

OBJECTIVE: Psychogenic non-epileptic seizures (PNES) are conversion disorders with functional neurological symptoms that can resemble epileptic seizures (ES). We conducted a systematic review to obtain an overview of the value of prolactin (PRL) levels in the differential diagnosis between PNES and ES. METHODS: We searched PubMed, EMBASE, and Cochrane Library databases for studies published up to June 4th, 2020. Published studies were included if they fulfilled the following criteria: original research on PRL changes after ES and PNES. By applying Bayes' theorem, we calculated the predicted values of PRL with pretest probabilities of 90 % and 75 % in ES. RESULTS: Sixteen studies were included in this review. All the studies showed that PRL levels increase after ES, especially 10-20 min after ES, when the elevation was most obvious. In studies where capillary PRL level measurements were included, the median sensitivity in the diagnosis of ES (all epileptic seizure types), generalized tonic clonic seizures (GTCS), focal impaired awareness seizures (FIAS), and focal aware seizures (FAS) was 67.3 %, 66.7 %, 33.9 %, and 11.1 %, respectively. The median specificity in the diagnosis of ES was 99.1 %. By using Bayes' theorem, when we used the median specificity and sensitivity for predictive value calculation, assuming a pretest probability of 90 %, a positive PRL measure was highly predictive (99 %) of all types of ES, and negative predictive values were all below 30 %. When we used the lowest specificity and sensitivity for predictive value calculation, assuming a pretest probability of 75 %, ES and GTCS had positive predictive values of 77.2 % and 81.0 %, respectively; the negative predictive values of PRL in ES and GTCS were 26.2 % and 29.6 %, respectively. CONCLUSIONS: The use of PRL could be a useful adjunct to differentiate GTCS from PNES. However, PRL levels are of limited use for differentiating FIAS or FAS from PNES, and a negative PRL measure is not predictive of PNES.

Induction of immune tolerance and altered cytokine expression in skin transplantation recipients.

Apoptotic lymphocytes can induce specific immune tolerance. This study aimed to investigate the influence of the preoperative transfusion of apoptotic lymphocytes on allograft survival after skin transplantation. In addition, we aimed to determine changes in IL-4, IL-10 and IFN-γ mRNA expression in the grafted skin. A total of 20 New Zealand white rabbits were randomly divided into two groups: lymphocyte preconditioned group (n = 10) and control group (n = 10). Rabbits in the lymphocyte preconditioned group were intravenously injected with 60Co γ-treated donor lymphocytes at seven days before the surgery. Rabbits in the control group were intravenously injected with normal lymphocytes at seven days before skin transplantation. The mRNA expression of IL-4, IL-10 and IFN-γ in the grafted skin was determined using real-time PCR. Skin allograft rejection time was 72.63 ± 2.65 days in the lymphocyte preconditioned group and 6.52 ± 0.64 days in the control group. IL-4, IL-10 and IFN-γ mRNA expression in the skin graft was 4.32 ± 0.48, 7.86 ± 0.56 and 2.63 ± 0.25 respectively in the lymphocyte preconditioned group, and 0.58 ± 0.07, 0.91 ± 0.14 and 8.68 ± 1.23 respectively in the control group. The preoperative transfusion of apoptotic lymphocytes induced immune tolerance in the grafted skin, as demonstrated by longer survival time of the grafts before rejection. This coincided with the increased mRNA expression of IL-4 and IL-10, and the decreased expression of IFN-γ.

[Study on Molecular Biology of Rare Two A307 Phenotypes].

OBJECTIVE: To identify the genotypes of the 2 blood samples whose serological typing were difficult by DNA sequencing analysis, and to investigate the molecular genetic basis of their genotypes. METHODS: The 2 blood samples were preliminary genotyped by PCR-SSP. The complete exon 6 and 7 in the ABO genes were amplified by PCR and the PCR products were directly sequenced and clonal sequenced in order to identify the genotypes. RESULTS: The forward typing showed that both samples were weak A, while the reverse typing showed that the samples contained anti-A1. They were preliminarily genotyped as A/O1. RESULTS: The sequencing analysis showed that the 2 samples contained the nt467C>T and nt745C>T mutation in the A allele, which resulted in an amino acid change from Proline (Pro) to Leucine (Leu) at codon 156 and also from Arginine (Arg) to Tryptophan (Trp) at codon 249. CONCLUSION: Through serology results and sequencing analysis, the 2 samples are identified as rare A307 phenotypes.