Objective:To investigate the role and possible mechanism of tripartite motif-containing protein 72 (Trim72) in acute viral myocarditis (AVMC) in mice.Methods:A mouse model of AVMC was established by intraperitoneal injection of Coxsackievirus B3 (CVB3, 2.0 × 10 5 PFU/mouse). Forty mice were randomly divided into the negative control (NC) + phosphate-buffered saline (PBS) group (NC+PBS group), Trim72 overexpression + PBS group (Trim72 + PBS group), NC + CVB3 group, and Trim72 + CVB3 group ( n = 10). Fourteen days before modeling, mice in each group were injected with adeno-associated virus type 9 vector (AAV9) encoding either negative control or Trim72 overexpression (5.0 × 10 11 VG/mouse) via tail vein. Subsequently, PBS or CVB3 was injected intraperitoneally in the PBS and CVB3 groups, respectively. After seven days, the surviving mice were euthanized, and the heart and serum samples were collected. HE and TUNEL staining were used to observe the cardiac pathological changes and cardiomyocyte apoptosis, respectively. The mRNA expression levels of Trim72 and pro-inflammatory cytokines TNF-α, IL-6, and IL-1β in myocardial tissues of each group were detected by RT-qPCR. The protein levels of cTnI, TNF-α, IL-6, and IL-1β in serum were detected by ELISA. The expression levels of Trim72, apoptosis-related proteins (Bax, Bcl-2, Cleaved caspase-3, Caspase-3), TLR4, p-p65, and p65 were detected by Western blot. Results:The protein and mRNA expression levels of Trim72 in myocardial tissues of mice in the NC+CVB3 group were significantly downregulated compared with those in the NC + PBS group ( P<0.05). Compared with the NC + CVB3 group, Trim72 overexpression significantly increased the protein and mRNA expression of Trim72 in myocardial tissues ( P<0.05), ameliorated myocardial inflammatory injury, decreased the apoptotic index of cardiomyocytes ( P<0.05), and reduced the levels of pro-inflammation cytokines TNF-α, IL-6, and IL-1β in the myocardium and serum ( P<0.05). Additionally, Trim72 overexpression also downregulated the protein expression of Bax, Cleaved caspase-3/Caspase-3, TLR4, and p-p65, and upregulated the protein expression of Bcl-2 in myocardial tissues ( P<0.05). There was no significant difference in the indexes of mice between the NC + PBS and Trim72 + PBS groups ( P>0.05). Conclusions:Trim72 overexpression attenuates AVMC in mice by inhibiting myocardial inflammatory injury and apoptotic imbalance, and the mechanism may be related to the negative regulation of the TLR4/NF-κB signaling pathway.

Objective:To explore the effects of calcium/calmodulin dependent protein kinase II (CaMKII) on myocardial ischemia-reperfusion injury in vitro, and apoptosis and autophagy of myocardial cells in isolated rats.Methods:Seventy female SD rats (250-280 g) with normal electrocardiogram were selected to establish the myocardial IR injury model by Langendorff perfusion system. These SD rats were randomly divided into five groups (n=14): cardiac ischemia reperfusion group (IR group), CaMKII phosphorylation activator group (IR+ isoproterenol group), CaMKII phosphorylation inhibitor analogue group (IR+KN92 group), CaMKII phosphorylation inhibitor group (IR+KN93 group), and control group. After reperfusion, the left ventricular function and myocardial morphology were measured to assess the myocardial injury, and TUNEL was performed to assess the apoptosis index. Western blot was used to determine the phosphorylation levels of CaMKII and PLN (p-CaMKII/CaMKII and p-PLN/PLN), and the expression levels of apoptosis-related proteins Bax, Bcl-2, cleaved caspase-3, and autophagy marker proteins LC3II/LC3I, Beclin-1 and P62.Results:Compared with the control group, the left ventricular function of the IR group was decreased, morphological arrangement of myocardial fibers was disordered, and the apoptosis index was increased. The levels of p-CaMKII/CaMKII, p-PLN/PLN, cleaved caspase-3, Bax/Bcl-2, LC3II/LC3I, and Beclin-1 were increased significantly, while the level of P62 was decreased significantly, and apoptosis and autophagy were increased significantly (all P<0.05). Compared with the IR group, the myocardial damage of rats in the IR+KN93 group was significantly improved, the apoptosis index was decreased, and the expression of p-CaMKII/CaMKII, p-PLN/PLN, Cleaved Caspase-3, Bax/Bcl-2, LC3II/LC3I and Beclin-1 were significantly decreased and the level of p62 was remarkable increased, and apoptosis and autophagy decreased significantly (all P< 0.05). Compared with the IR group, the left ventricular function was further decreased in the IR+ isoproterenol group, while the levels of apoptosis and autophagy were further increased ( P < 0.05), while there was no significant difference in myocardial indexes between the IR+ KN92 group and the IR group ( P > 0.05). Conclusions:Inhibition of CaMKII phosphorylation attenuates isolated myocardial ischemia-reperfusion injury by reducing apoptosis and autophagy.

Objective:To investigate the effect of prenatal taurine supplementation on sensorimotor ability and synaptophysin (Syn) expression in the hippocampus of juvenile rats with intrauterine growth restriction (IUGR).Methods:The IUGR rat model was induced by food restriction throughout pregnancy.Pregnant rats were randomly divided into normal control group, IUGR group and IUGR+ taurine group.Sensorimotor ability was tested in 2-week-old juvenile rats via grading the tail suspension scores and beam balance test scores, followed by detecting Syn expression in the hippocampus of juvenile rats by immunohistochemistry and Western blot.The correlation between sensorimotor ability scores and Syn expression was assessed.Results:Tail suspension time[(14.62±3.46) s vs.(25.38±5.92) s, P<0.001] and beam balance test scores [(9.08±1.38) scores vs.(12.08±1.16) scores, P<0.001] in the IUGR group were significant lower than those of normal control group.Tail suspension time (22.77±5.16) s and beam balance test scores (11.08±1.38) scores in IUGR+ taurine group were significantly higher than those in IUGR group (all P<0.05), but there was no significant difference comparable to those in normal control group ( P>0.05). The average optical density ( A) value [(53.96±2.37)% vs.(61.68±3.07)%, P<0.001] and protein expression of Syn (1.82±0.23 vs.2.23±0.17, P<0.001) in rat hippocampus of IUGR group were all signi-ficantly lower than those in normal control group.The A value [(60.27±2.59)%] and expression of Syn protein (2.07±0.17) in IUGR+ taurine group were significantly higher than those in IUGR group (all P<0.05), but there was no significant difference comparable to those in normal control group ( P>0.05). The expression of Syn in rat hippocampus was positively correlated with the tail suspension test time and beam balance test scores (all P<0.05). Conclusions:Prenatal taurine supplementation can improve the sensorimotor ability of juvenile rats with IUGR by upregulating Syn in the hippocampus.

AIM To establish an ultra-performance liquid-chromatography tandem mass spectrometry (UPLC-MS/MS) method for the simultaneous content determination of five constituents in Jiangzhi Huoxue Tablets (Polygoni multiflori Radix,Astragali Radix,Chuanxiong Rhizoma,etc.).METHODS The 50％ methanol extract of this drug was performed on a 40 ℃ thermostatic Restek UItra BiPh column (100 mm × 2.1 mm,5 μm),with the mobile phase comprising of acetonitrile (containing 0.1％ formic acid)-0.1％ formic acid flowing at 0.4 mL/min in a gradient elution manner.RESULTS Stilbene glycoside,tanshinone Ⅱ A,emodin,ferulic acid and puerarin showed good linear relationships within the ranges of 4.01-1 027,0.7-187,1.48-380,3.98-1 020 and4.285-1 097 ng/mL (r ＞0.994 0),whose average recoveries were 98.57％-101.0％ with the RSDs (n =6) of less than 4.79％.CONCLUSION This specific and sensitive method can be used for the quality control of Jiangzhi Huoxue Tablets.

Objective To evaluate the clinical superiority of transumbilical mini pure single-port laparoscopic Palomo's surgery on the treatment of primary teenage varicocele.Methods All operations were successfully completed using mini pure single-port laparoscopic Palomo's surgery.Its clinical effects were evaluated by the operative time,blood loss volume,intra-or post-operative complications,and hospital stay.Six to eighteen months after surgery,follow-up was made in 78 patients.Results All the operations were successful.The operation time was 5-15 (mean 8) minutes,the intra-operative loss blood was 1-5 (mean 1.5) mL,and the post-operative hospital stay was 2-4 (mean 3) days.No intra-operative or post-operative complications happened.Sperm analysis of 95％ patients came back to normal in 6-18 months.Conclusions The transumbilical mini pure single-port laparoscopic Palomo's technique has more obvious superiority and it is worthy of clinical application and generalized widely.

OBJECTIVETo provide a new therapeutic approach for Staphylococcus epidermidis biofilm-associated infections by the study of inhibitory effect of andrographolide (AG) on S. epidermidis biofilm.

METHODS. epidermidis biofilms were set up in vitro, erythromycin was acted as the positive control agent, XTT reduction assay was used to evaluate AG on the initial adhesion of S. epidermidis and bacterial metabolism within biofilm, microscope was applied to observe biofilm morphology, and Congo red assay was used to detect polysacchatide interc-ellular adhesion (PIA)formation when exposed to AG.

RESULTAG showed inhibitory effects against the initial adhesion of S. epidermidis at concentrations of 1 000,100, 10 mg x L(-1), respectively,and inhibited metabolism of biofilm bacteria at the concentration of 31.25 mg x L(-1), and exhibited significantly inhibition against the biofilm morphology at the concentration of 250 mg x L(-1), while did not display inhibition against PIA formation at the concentration of 10 mg x L(-1).

CONCLUSIONAG could remarkably inhibit biofilm formation of S. epidermidis, although it was less potent than erythromycin.

Bacterial Adhesion ; drug effects ; Biofilms ; drug effects ; Diterpenes ; pharmacology ; Dose-Response Relationship, Drug ; Erythromycin ; pharmacology ; Staphylococcus epidermidis ; drug effects ; physiology