Homocysteine ; pharmacology ; Human Umbilical Vein Endothelial Cells ; drug effects ; metabolism ; Humans ; Lipid Peroxidation ; drug effects ; Metallothionein ; pharmacology

Cardiovascular Diseases ; Growth Differentiation Factor 15 ; Humans

Abstract: Objective　To explore the antiviral effect of baricitinib in the SARS-CoV-2 infection and influence on cytokine levels. Methods　Calu-3 cells were infected with SARS-CoV-2 at MOI of 0.1, and the levels of inflammatory cytokines (IL-6, IL-8, TNF-α and IL-1β), interferon β (IFN-β) and interferon-stimulated gene, IFIT2 in the infected cells were analyzed by qRT-PCR methods. At the same time, Calu-3 cells were infected with SARS-CoV-2 (MOI=0.1) after being treated with baricitinib for 2 hours. Cells were collected at 0, 24, 36, and 48 hours, and analyzed for the mRNA of the above genes in the drug-treated and untreated groups. Results　The mRNA levels of IL-6, TNF-a, IL-1β, IFN-β and IFIT2 in Calu-3 infected by SARS-CoV-2 cells were increased significantly. These cytokines were increased by nearly 100-fold post-infection 48 h compared with the control (P<0.000 1), and continued to increase with the infection time (P<0.001 or P<0.000 1). The increase of IL-8 mRNA level was not as significant as IL-6, TNF-α, IL-8, IL-1β, but it also showed a 2-4 folds increase. Baricitinib does not affect the level of viral RNA in Calu-3 cells after SARS-CoV-2 infection (P>0.05). However, baricitinib can significantly inhibit the up-regulation of IL-6 and TNF-α levels induced by SARS-CoV-2 infection (5.25-fold and 3.90-fold down-regulation, respectively, P<0.01), and has little effect on the levels of IL-8 and IL-1β . In addition, the drug could also significantly down-regulate the increase in IFN-β and IFIT2 levels caused by viral infection (10.51-fold and 90.78-fold down-regulation, respectively, P<0.000 1). Conclusions　Baricitinib inhibits the release of inflammatory cytokines to some extent, but it drastically down-regulates the expression of interferons and interferon-stimulated genes (ISGs), and has limited antiviral effect on SARS-CoV-2. Considering that interferon signal pathways play important roles on viral infection, caution should be exercised when using baricitinib to treat COVID-19 patients.

OBJECTIVETo observe the clinical therapeutic effect of Nuzhen Yunyu Decoction (NYD, a Chinese recipe for nourishing blood and reinforcing Shen) and its influence on the hemodynamic parameters in ovarian and uterine arteries.

METHODSSixty-eight patients were randomly divided into two groups at the ratio of 2:1, 45 in the TCM group treated with Chinese medicine and 23 in the control group treated with clomiphene citrate. Menstrual cycle were taken as the indexes for clinical observation; basal body temperature, cervical mucus, vaginal exfoliative cells, serum hormones and B ultrasonic examination were taken as the indexes for ovulation and pregnancy assessment. The hemodynamic parameters of uterus and ovary were determined by using color Doppler ultrasonography, and the growth of follicles and ovulation, thickness of uterine endometrium were also monitored continuously.

RESULTSResults of 3 cycles treatment and 1 year follow-up study showed that the ovulation promoting rate, pregnancy rate and abortion rate in the two groups were similar, but the treated group showed better effects than that of control group, in such aspects as regulating menstruation, promoting growth and development of follicle, strengthening endometrium, improving blood supply and circulation of uterine and ovary (P < 0.05, P < 0.01).

CONCLUSIONNYD has the effects of ovulation promotion and holistic regulation of reproductive system.

Adult ; Anovulation ; drug therapy ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Hemodynamics ; Humans ; Infertility, Female ; drug therapy ; etiology ; Ovary ; blood supply ; Ovulation Induction ; Phytotherapy ; Uterus ; blood supply ; Yin Deficiency ; drug therapy

The rate of corneal graft rejection is still high for high-risk keratoplasty although immune suppression drug is routinely used.The role of traditional Chinese medicine in corneal transplantation is concerned gradually.Heijingpaichitang on the prevention and treatment of rats with high-risk corneal allograft rejection needs further study.Objective This study was to investigate the inhibitory effect of heijingpaichitang on high-risk corneal transplantation immune rejection in rats.Methods Sixteen female SD rats were used as the donors and 32female Wistar rats were served as recipients.The high-risk corneal trasplantation models were established by corneal suture in 32 Wistar rats,and then homogeneity variant SD-Wistar corneal transplantation was performed.The recipients were randomized into model control group,cyclosporinc A (CsA)group,heijingpaichitang group and CsA +heijingpaichitang group.CsA,heijingpaichitang and CsA + heijingpaichitang was orally administered 4 days after operation once per day for 15 days,and normal saline solution was used at the same way in the model control group.Ocular anterior segment reaction was examined under the slit lamp and corneal opacification,edema and neovasculation were scored based on Larkin' s criteria.Rejection index of the corneal graft was recorded and the graft survival time was calculated.The pathological examination of the corneal graft was carried out in all rats,and the inflammatory cells in the corneas and CD4+ cells in the periphery blood were assayed using flow cytometry.The use of the animals complied with ARVO Statement.Results Corneal graft rejection occurred in 10 days after operation in the model control group,12-13 days in the CsA group and heijingpaichitang group and 22 days in the CsA +heijingpaichitang group.Compared with model control group,the scores of the corneal opacification,corneal edema and neovascularization were significantly lower in the CsA group,heijingpaichitang group and CsA+heijingpaichitang group (P＜0.05),and all the scores were declined in the CsA+ heijingpaichitang group compared with CsA group and heijingpaichitang group(P＜0.01),but no significant differences were seen in the scores between the CsA group and heijingpaichitang group(P＞0.05).The mean survival time of grafts was (10.38 ±1.69)days in the model control group,(22.50 ± 3.07) days in the CsA + heijingpaichitang group,with the significant difference (t =-9.790,P =0.000).The pathological examination of graft showed that the lymphocytes and new blood vessels were less in the CsA+heijingpaichitang group compared with CsA group and heijingpaichitang group 15 days after operation.Flow cytometry verified that the number of lymphocytes in graft,CD4+cells and CD4+/CD8+ in periphery blood were significantly lower in the heijingpaichitang group,CsA group and CsA+heijingpaichitang group compared with model control group (P＜0.05).Conclusions Heijingpaichitang can inhibit immune rejection to certain extent in high-risk corneal transplantation rat and has a similar effect to 0.1％ CsA.Heijingpaichitang and 0.1％ CsA have a synergistic effect.

Objective To establish an ultraviolet-irradiation damage model in cultured fibroblasts derived from human skin and to explore the potential protective effects and mechanisms of amyloid precursor protein 17-met peptide (APP17-mer peptide) on the oxidative damage and collagen metabolism in cultured fibroblasts after ultraviolet irradiation. Methods Human dermal fibroblast cultures were established by outgrowth from foreskin biopsies of a healthy donor and were irradiated by a single exposure to ultraviolet rays and cultured in a series of concentrations of APP17-mer peptide (0, 20, 40, 80 μmol/L).The activity of fibroblasts was detected by the assay of MTT. The intracellular ROS level was measured with a confocal microscope. The expression of MMP-1 mRNA was analyzed real-time quantitatively following RT-PCR. Results Primary cultures of human skin fibroblasts were established from human foreskin in DMEM supplemented with 10 % fetal bovine serum. UV irradiation depressed cellular activity and increased intracellular level of ROS (P<0.05). 40μmol/L and 80μmol/L APP17-mer peptide increased the cellular activity in both UV irradiated fibroblasts and unirradiated fibroblasts (P<0.05), however,20 μmol/L did not show such protective effects (P>0. 05). 40μmol/L APP17-mer peptide could depress the level of ROS in irradiated libroblasts. A single exposure of fibroblasts to UV irradiation resulted in 1.78 foldup-regulation of MMP-1 mRNA compared with unirradiated sample, 40μmol/L and 80μmol/L APP17-mer peptide decreased the expression of MMP-1 mRNA (P<0.05 and P<0.01, respectively).Conclusion APP17-mer peptide can enhance cellular activity under UV-induced oxidative stress and in-hibit collagen degradation in fibroblasts irradiated with ultraviolet rays. Inhibition of ROS production may be involved in the protective mechanism of APP17 peptide.

AIMOn the model of limb ischemia/reperfusion (LIR), the effects of taurine on pulmonary morphological changes in rats were observed.

METHODSWistar rats were divided into three groups (n=8): control group, ischemia/reperfusion group (IR) and taurine + IR (Tau + IR). Then macroscopic inspection and optical and transmission electron microscopies (TEM) were performed to assess the morphological changes of the lung tissues and their lung index (LI) and lung permeability index (LPI) and reactive oxygen species (ROS), malondialdehyde (MDA) were measured as well.

RESULTSThe morphological changes of lung tissue after LIR were characterized by an increase of permeability of the alveoli-capillary membrane and infiltration of inflammatory cells. Under optical microscopy, there were congestion and swelling in pulmonary microvessels with broadened the spaces around the blood vessels. Under TEM, a number of tight-junctional regions between adjacent alveolar epithelial cells and between pulmonary microvessels endothelium were "open". The LI, LPI, MDA and ROS increased. The specimens of Taurine + IR group revealed slight to moderate degrees of damages in the lung tissues.

CONCLUSIONTaurine protects lung from LIR in rats and the protective action on tight-junctional regions between cells and anti-oxygen is one of the protective mechanisms of taurine.

Animals ; Hindlimb ; blood supply ; Lung ; drug effects ; pathology ; Lung Injury ; prevention & control ; Male ; Malondialdehyde ; analysis ; Rats ; Rats, Wistar ; Reactive Oxygen Species ; analysis ; Reperfusion Injury ; prevention & control ; Taurine ; pharmacology ; Tight Junctions ; metabolism

Objective To investigate the expression of caspase-10 in differentiated thyroid carcinoma and association with its development and metastasis. Methods Thyroid samples from 37 patients in a period from January 2006 to December 2007, with differentiated thyroid carcinoma were retrospectively analyzed for caspase-10 by immunohistocbemistry(streptavidin-perosidase, S-P), compared to control group of 46 cases with nodtdar goiter. The relationship between the expression of caspase-10 and the clinical pathologic characteristics of thyroid carcinoma were also explored simultaneously. Results caspase-10 were observed as brown or yellow particles located in the cytoplasm or cell membrane of nodular goiter but there were no significant evidence for its positive expression in thyroid carcinoma, caspase-10 expression was markedly down-regulated in differentiated thyroid carcinoma(29.73%,11/37) compared with benign nodules(71.74%,33/46, χ2=14.528, P<0.01). The positive expression in 18 cases with lymph node metastasis(11.11%,2/18) was significantly lower than those in 19 patients without lymph node metastasis(47.37%,9/19; χ2=4.210, P<0.01). There was no significant correlation(P> 0.05) between the expression of caspase-10 and the clinical pathologic characteristics including male, age, TNM stage and pathologic type. Conclusion Down-regulation of caspase-10 may play a critical role in carcinogenesis and development of differentiated thyroid carcinoma.

Objective:To investigate the effect of paraformaldehyde fixation on measuring the protein-protein interaction by fluorescence resonance energy transfer(FRET)to resolve the problem of FRET efficiency calculation in excess-movement cells.Methods:The C terminals of TCR ? chain(TRA)and TCR ? chain(TRB)genes,which were ideal for protein-protein interaction research,were fused with ECFP and EYFP gene respectively by fusion PCR and transferred into target cell.A grou Pcells were fixed in paraformaldehyde(0.5%)for 0.5~1h and another left alive,then these cells were subject to ECFP/EYFP FRET calculation with confocal laser scanning microscope.The ECFP/EYFP FRET efficiencies in live and fixed cell were analyzed and compared.Results:There is no significant statistical difference between the ECFP/EYFP FRET efficiencies of live cell and cell fixed with lower paraformaldehyde concentration and shorter incubation time.Conclusion:fixation with low-concentration paraformaldehyde and short-time incubation has no distinct influence on measuring protein-protein interaction,and facilitated the FRET calculation in excess-movement cells.

Objective To establish a method of detecting hepatitis B virus x antigen (HBxAg) and antibody to HBxAg (anti-HBx) and to demonstrate its clinical significance of HBxAg and anti-HBx in sera from patients of chronic hepatitis B (CHB),liver cirrhosis (LC) and hepatocellular carcinoma (HCC). Methods Full length HBx gene was cloned into pET30a(+),a prokaryotic expression vector,named pET30a-X.It was transformed into Escherichia coli BL21 (DE3),followed the fusion protein of HBx-His was induced by IPTG.The purified fusion protein was used to immunize rabbit as an antigen to generate polyclonal antibody to HBx protein.The method of enzyme-linked immunosorbent assay (ELISA) was established by using purified fusion protein and generated antibody,which was used to detect HBxAg and anti-HBx in sera from patients of CHB,LC,HCC and normal healthy people.Results The positive rates of HBxAg/anti-HBx were 8.7%/10.4% for CHB,17.9%/40.6% for LC,and 9.8%/34.4% for HCC, respectively.In statistics,the positive rates of anti-HBx in LC and HCC were higher than that in CHB (P