[Cryptotanshinone May Induce Ferroptosis of Human Liver Cancer HepG2 Cells].

Objective To observe the effect of cryptotanshinone on the ferroptosis of human liver cancer HepG2 cells. Methods The viability of the HepG2 cells cultured in vitro was determined using the Cell Counting Kit-8(CCK-8),and the half maximal inhibitory concentration(IC50)was calculated.The cell morphology was observed using an inverted microscope.The reactive oxygen species(ROS)level was detected with the 2',7'-dichlorodihydrofluorescein diacetate(DCFH-DA)probe.The glutathione(GSH)assay kit was used to determine the GSH level.Western blot analysis was employed to detect the expression of cystine/glutamate antiporter system light chain(xCT)and glutathione peroxidase 4(GPX4),two marker proteins in ferroptosis.Additionally,the cell viability,ROS level,GSH level,and the expression levels of xCT and GPX4 were detected for the cells treated with the ferroptosis inhibitor ferrostain-1(Fer-1),the iron chelator deferoxamine(DFO),and the ROS scavenger N-acetylcysteine(NAC).Results Cryptotanshinone significantly inhibited the cell viability of HepG2 cells with an IC50 of 93.73 µmol/L,and caused the morphological changes and death of the cells.It could significantly induce ROS accumulation,reduce GSH level,and down-regulate the expression of xCT and GPX4 in HepG2 cells.Fer-1,DFO,and NAC can remedy the cryptotanshinone-caused decrease in the cell viability of HepG2 cells.Fer-1 could inhibit cryptotanshinone-induced ROS accumulation,restore GSH level,and recover the expression of xCT and GPX4. Conclusion Cryptotanshinone may increase the accumulation of ROS by inhibiting the expression of xCT and GPX4 to induce the ferroptosis of HepG2 cells.

The Role of the NOD1/Rip2 Signaling Pathway in Myocardial Remodeling in Spontaneously Hypertensive Rats.

BACKGROUND Chronic hypertension changes the function and structure of the heart and blood vessels. This study aimed to explore the role of the NOD1/Rip2 (nucleotide-binding oligomerization domain 1/receptor-interacting protein 2) signaling pathway in myocardial remodeling in spontaneously hypertensive rats (SHRs). MATERIAL AND METHODS Blood pressure was measured using a tail cuff. The cardiac structure was observed using echocardiography. Slices of the myocardium were stained with hematoxylin and eosin. The expression of NOD1 and Rip2 was detected using real-time polymerase chain reaction, western blot, and immunohistochemistry. The content and distribution of collagen in the myocardium were observed using Van Gieson staining. Enzyme-linked immunosorbent assay was used to detect the interleukin-1 (IL-1) concentrations. SHRs were treated with the NOD1 agonist iE-DAP and NOD1 inhibitor ML130. RESULTS The NOD1 agonist increased blood pressure in SHRs, and the NOD1 inhibitor decreased blood pressure; the interventricular septum thickness (IVST) and left ventricular posterior wall thickness (LVPWT) of the agonist-treated group were thicker than those of the control group, and the antagonist exerted the opposite effects. The levels of the NOD1 and Rip2 mRNAs and proteins, serum IL-1 concentration, and myocardial collagen volume fraction (CVF%) increased in SHRs in the NOD1 agonist group, but the levels of NOD1 and Rip2, serum IL-1 concentration, and myocardial collagen volume fraction (CVF%) decreased in SHRs in the NOD1 inhibitor group. CONCLUSIONS NOD1/Rip2 expression increased during the progression of myocardial remodeling in SHRs. The NOD1 agonist increased NOD1 expression and promoted myocardial remodeling, while the NOD1 antagonist reduced NOD1/Rip2 expression and protected against myocardial remodeling.

[Antiasthmatic effects of different tonifying kidney-Yin formulas and their effects on airway remodeling in chronic asthma].

Both of Zuogui Wan(ZGW) and Liuwei Dihuang Wan(LWDHW) contain ingredients of Sanbufang(SBF), which have been proven to have antiasthmatic effects. In order to study the antiasthmatic effects of the three tonifying kidney-Yin formulas and their mechanisms, BALB/c mice were randomly divided into 5 groups. Chronic asthma was induced by ovalbumin. Mice in treated groups were respectively given 49.0 g•kg⁻¹ZGW, 35.0 g•kg⁻¹LWDHW and 22.4 g•kg⁻¹SBF by gavage. Those in normal and model group were given normal saline. After treatment, sneeze and nose scratching times of mice were observed. Histological lung sections were prepared to determine the basement membrane thickness(BMT), smooth muscle thickness(SMT), collagen area(CA) and numbers of goblet cells(GCN). Western blotting and RT-PCR were used to determine the expression levels of MMP-9, TGF-ß1, Smad2, Smad3 and Smad7. The results showed that sneeze and nose scratching times of ZGW group were significantly lower than those of SBF group. Its inhibition degree on airway remodeling was significantly higher than SBF group. Sneeze and nose scratching times of LWDHW group were significantly lower than SBF group. Its CA and GCN were significantly lower than SBF group. Regarding the four airway remodeling related factors, MMP-9, TGF-ß1, Smad2 and Smad3 of ZGW group were significantly lower than those of SBF group, and its Smad7 was significantly higher than SBF. Smad7 of LWDHW group was significantly higher than SBF. There was no significant difference in MMP-9 between model group and SBF group. The results indicate that there are significant differences in the antiasthma effect of these tonifying kidney-Yin formulas. The regulatory effects of ZGW and LWDHW on MMP-9 and Smad7 may be correlated with the differences in the inhibitory effect of airway remodeling of the three formulas.

[Effect of Yupingfeng granule on cytokines of allergic rhinitis induced by OVA in rats].

To investigate the effects of Yupingfeng granule (YPF) on immune factors of the rats with allergic rhinitis (AR) induced by ovalbumin(OVA). OVA 0.3 mg, Al(OH)3 30 mg and saline 1 mL were mixed and intraperitoneally injected for the initial immunization, 4% OVA 200 µg (50 µL) was given to the nose on the 15th day for the second immunization to establish the allergic rhinitis model. Sixty male SD rats were randomly divided into allergic rhinitis(AR) model group, Yupingfeng granule three dose (2.7,1.35,0.68 g•kg⁻¹) groups, control drug Biyankang (0.4 g•kg⁻¹) and normal control group. After 14 days, efforts were made to collect blood from abdominal aorta, and take nasopharynx tissues and fasten them into 10% formaldehyde for a pathological examination. The levels of HIS, IgE, IL-4 and TNF-α in serum were examined by radioimmunoassay, and nasal mucosa tissues were examined by HE staining. According to the results, the levels of HIS, IgE, IL-4 and TNF-α in serum of Yupingfeng granule groups were significantly lower than that of AR model group (P<0.05, P<0.01). Nasal mucosa tissues showed slight morphological changes and inflammatory cell infiltration, with unobvious necrosis. Yupingfeng granule can improve the pathological changes of nasal mucosa tissues, and reduce the production and release of immune factors during allergic rhinitis (AR) process in vivo by OVA, which may be the important curative mechanism of allergic rhinitis.

Integrated coagulation-trickling filter-ultrafiltration processes for domestic wastewater treatment and reclamation.

More and more research effort has been put into the development of affordable and high-efficiency wastewater reclamation technology for small communities. In this study, an integrated chemically enhanced primary treatment (CEPT), trickling filter (TF) and ultrafiltration (UF) process was developed with success. Coagulant produced from fly ash was used to enhance primary treatment, while trickling filter packed with coal cinder through four-layer structure without aeration was employed for further removal of COD and ammonium-nitrogen from the CEPT effluent. 95 and 88% removal of COD and ammonium were achieved, while total phosphorus (TP) and suspended solid (SS) were found to be removed completely at a coagulant dosage of 2.5 mL/L in the CEPT-TF-UF system. The product water can meet the standard of Reuse of Recycling Water for Urban Water Quality Standard for Urban Miscellaneous Water Consumption (GB/T 18920-2002, China).

Indigoferavallicola (Fabaceae), a new species from Yunnan, southwest China.

Indigoferavallicola (Fabaceae), a new species is described and illustrated. This plant is only found from two localities in the central Yunnan Province, southwest China. It is characterized by having the prostrate habit, usually 13-17-foliolate leaves and the relatively small (3-5 mm long) flowers. Morphological comparisons with its closest relatives, I.rigioclada, I.franchetii, I.chaetodonta, and I.henryi are also presented.

Silenevanchingshanensis (Caryophyllaceae) a new species from Southwest China.

Silenevanchingshanensis (Caryophyllaceae), a new species from Fanjingshan Mountain in Guizhou (southwest China) is described and illustrated. It is morphologically similar to S.morrisonmontana and S.hupehensis, from which it can be easily distinguished by having pubescent stems usually 10-15 cm long, linear-oblanceolate leaves 3-6 cm × 3-6 mm, often 2-5-flowered cymes, pink or violet petals and narrowly ovoid capsules.

[Diagnostic significance of NK cell activity in the patients with hemophagocytic syndrome].

This study was aimed to explore the level of NK cell activity in the patients with secondary hemophagocytic syndrome (HPS) and its significance for early diagnosis of this disease. 16 suspected HPS patients and 25 healthy subjects were enrolled in this study. The activity of NK cells in peripheral blood was detected by a released LDH assay. The activity level of NK cells in peripheral blood from patients who were finally diagnosed as HPS was compared with healthy subjects. The results showed that 8 out of 16 suspected HPS patients were finally diagnosed as secondary HPS. The activity of NK cells in peripheral blood of these 8 patients was obviously lower than that in healthy subjects with statistical significance (p<0.001), and showed abnormal in the early stage of this disease. It is concluded that the detection of NK cell activity may play an important role in earlier diagnosis of secondary hemophagocytic syndrome.

[Transforming growth factor beta alleviates acute graft-versus-host-disease after allogeneic bone marrow transplantation in murine model].

This study was purposed to investigate the effects and mechanism of transforming growth factor beta (TGF-beta) on acute graft-versus-host disease (aGVHD) after allogeneic bone marrow transplantation (allo-BMT). The recipients were male BABL/c mice, while the donors were male C57BL/6 mice. The murine model of aGVHD had been established by allo-BMT with donor derived T cells. Experiment was divided into four groups: control group, radiation control group, transplantation control group and TGF-beta treated group. Mice in TGF-beta treated group were daily subcutaneously injected TGF-beta1 (1 microg/kg) in two days before transplantation until seven days after it. The results showed that the survival time of mice in TGF-beta treated group was significantly longer than that in transplantation control group, and the aGVHD pathological changes in TGF-beta treated group were milder than that in transplantation control group. At seven days after transplantation, the level of IL-2 in TGF-beta treated group was significantly higher than that in control group, but significantly lower than that in transplantation control group. The level of IL-10 in TGF-beta treated group was significantly higher than that in transplantation control group, but the level of IL-10 in transplantation control group was significantly lower than that in other groups. It is concluded that TGF-beta may alleviate or suppress lethal aGVHD, and elevate the survival rate after allo-BMT in murine model. Accommodating of the Th1 and Th2 cytokine levels is the possible mechanism of TGF-beta preventing lethal aGVHD.

The porcine alpha1, 3 galactosyltransferase gene siRNA targeted heterozygous hepatocyte negative express GT / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To study whether the porcine alpha1, 3 galactosyltransferase gene siRNA targeted heterozygous hepatocyte negatively expresses GT mRNA and resists to the cytotoxicity of nature antibody in human serum.</p><p><b>METHODS</b>The porcine alpha1, 3 galactosyltransferase gene siRNA targeted vector (pPNTloxPGTsiRNA) were construct with pPNTloxPGT and pMXSV/U6 vector. Positive-negative selection was used to produce a heterozygous pPNTloxPGTsiRNA knockout (+/-) clone. The GT mRNA expressions were detected with northern blot. Complement-mediated NAb cytotoxicity after incubation of hepatocytes with NAbs and complement was determined using 3- (4, 5-dimethylthiazol-2-yl)-5-(3-carboxymethyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium(MTS, tetrazolium salt) colorimetric assay.</p><p><b>RESULTS</b>The pPNTloxPGTsiRNA targeted porcine hepatocyte (+/-) negative express GT mRNA. Only 14% to 18% cytotoxicity can be detected at the highest serum concentration. The pPNTloxPGT targeted porcine hepatocyte (+/-) express GT mRNA just as the wild type porcine cells and the cytotoxicity are 77% to 83%.</p><p><b>CONCLUSION</b>The porcine a1, 3 galactosyltransferase gene siRNA targeted heterozygous hepatocyte (+/-) negative express GT and resisted to nature antibody in human serum.</p>