Objective To explore the angiogenic function of EPC from peripheral blood in kidney transplanted patient and to reveal its regulative mechanism.Methods 23 chronic renal failure patients without diabetes were recruited in department of Urology Peking University Third Hospital from January 2014 to February 2015.Fasting peripheral blood mixed with heparin (20 U/mL) was collected one day before and 24 hours after kidney transplantation.We set preoperative blood as control and the postoperative blood as the experimental group.EPC from peripheral blood were isolated by density-gradient centrifugation.FACS was used to identify the EPC.The AA metabolites PGE2 in EPC cultured medium was measured by liquid chromatography-tandem mass spectrometry (LC-MS/MS).Q-PCR and WB were used to detect the expression of endothelial markers in HUVEC cultured under the EPC conditional medium.Tube formation assay was performed to assess the angiogenic ability of HUVEC.Results EPC from kidney transplantation expressed c-kit and CD31 by FACS analysis.Multiple types of AA metabolites was detected in the conditional medium by LC-MS/MS and level of PGE2 increased into two folds after kidney transplantation, compared with that before operation(P ＜ 0.05).HUVEC highly expressed CD31 and VE-cadherin cultured under conditional medium, which were 1.5 folds compared with that before operation (P ＜ 0.01).And those cells formed more tubes than that in control group, which showed better angiogenic capacity.HUVEC, treated by PGE2, had the similar biological characteristics like the conditional culture.Conclusions EPCs in the peripheral blood form kidney transplantation patient secret the PGE2, which can enhance the capacity of angiogenesis in HUVEC.

Herpesviruses is one of the most common human infectious diseases, which can be divided into different types based on clinical infection degree.Herpes simplex virus usually results in buccal and genital mucocutaneous infections, while cytomegalovirus is the most common opportunistic pathogen associated with significant morbidity and mortality in immunocompromised hosts, especially in transplant and cancer patients.Although nucleoside analogues are effective antiviral drugs, the emergence of drug-resistant viruses has created a barrier for the treatment of herpesviruses infections, especially in immunocompromised patients.Therefore, novel therapeutic agents are needed to avoid the limitations of drug resistance.In this article, research progress in the therapeutic agents for drug-resistant herpesviruses was reviewed from the aspects of non-nucleoside analogues, novel antiviral targets and newly antiviral mechanisms.

OBJECTIVE:To establish a method for content determination of steroidal saponin in Allium sativum. METHODS:Pre-column derivatization of steroidal saponin was performed by using the derivatization agent of nitro-benzoic acid-chlorine. And HPLC was conducted to determine the content of steroidal saponin. The column was Shimadzu VP-ODS with mobile phase of aceto-nitrile-water mixed solution(80:20,V/V)at a flow rate of 1.0 ml/min,the detection wavelength was 254 nm,the column temper-ature was 25 ℃,and the injection volume of 20 μl. RESULTS:The linear rang of sarsasapogenin was 0-1.25 mg/ml(r=0.999 0);RSDs of precision,stability and reproducibility tests were lower than 2%;recovery was 93.1%-96.8%(RSD=1.56%,n=6). CON-CLUSIONS:The method is simple,stable with good separation,and can be use for the content determination of steroidal saponin in A. sativum.

OBJECTIVETo observe the effects of electroacupuncture (EA) at different acupoints on apoptosis-related serum and expression of microRNA (miRNA) in rats with myocardial ischemia, so as to explore its mechanism of action.

METHODSA total of 48 male Wistar rats were randomly divided into a normal group, a model group, a Neiguan group and a acupoint compatibility group, 12 rats in each group. Isoprenaline hydrochloride (ISO) with a daily dose of 2 mg/kg was subcutaneously injected for 14 days to establish the myocardial ischemia model in the model group, Neiguan group and acupoint compatibility group. Rats in the normal group were subcutaneously injected with an equal volume of normal saline. After modeling, rats in the Neiguan group were treated with EA at "Neiguan" (PC 6), while rats in the acupoint compatibility group were treated with EA at "Guanyuan" (CV 4), "Zusanli" (ST 36) and "Neiguan" (PC 6). Rats in the normal group and model group were treated with immobilization, once day for 21 days. The contents of creatine kinase-MB (CK-MB), vascular cell adhesion molecule-1 (VCAM-1) and endothelin-1 (ET-1) in serum were detected by enzyme-linked immunosorbent assay (ELISA); apoptosis index (AI) of myocardial cells was detected by TUNEL method; the expressions of miRNA-1, miRNA-133, miRNA-208 and miRNA-499 were detected by real-time PCR method.

RESULTSCompared with the normal group, the serum CK-MB, VCAM-1 and ET-1 were significantly increased in the model group, Neiguan group and acupoint compatibility group (all P < 0.01), and the apoptosis index was significantly increased (all P < 0.01). The CK-MB, VCAM-1 and ET-1 in the Neiguan group and acupoint compatibility group were significantly lower than those in the model group (all P < 0.01); the AI was reduced, which was more significant in the acupoint compatibility group (P < 0.05). Compared with the normal group, the expression of miRNA-133 was reduced (P < 0.01) and those of miRNA-208, miRNA-1 and miRNA-499 were significantly increased in the model group (all P < 0.01). Compared with the model group, the expression of miRNA-133 was increased (both P < 0.01) and that of miRNA-208, miRNA-1 and miRNA-499 were significantly reduced (all P < 0.01) in the Neiguan group and acupoint compatibility group. Compared with the Neiguan group, the expression of miRNA-133 was increased (P < 0.01) and those of miRNA-208, miRNA-1 and miRNA-499 were significantly reduced in the acupoint compatibility group (P < 0.01, P < 0.05).

CONCLUSIONEA at acupoints, especially acupoint compatibility group, could effectively prevent and treat myocardial ischemia, and the protective effect is possibly correlated to the double regulation on increasing the expression of miRNA-133 and inhibiting the expression of miRNA-1, miRNA-208, miRNA-499.

Acupuncture Points ; Animals ; Apoptosis ; Disease Models, Animal ; Electroacupuncture ; Endothelin-1 ; genetics ; metabolism ; Humans ; Male ; MicroRNAs ; genetics ; metabolism ; Myocardial Ischemia ; genetics ; metabolism ; physiopathology ; therapy ; Myocytes, Cardiac ; metabolism ; Rats ; Rats, Wistar

BACKGROUND: Imatinib has a significant pro-apoptosis effect on chronic myelogenous leukemia (CML), but there are still some patients being resistant to it. Human umbilical cord mesenchymal stem cells (hUC-MSCs) affect the apoptosis of a variety of hematologic malignancies. However, the impacts of hUC-MSCs on the apoptosis of CML cells induced by imatinib remain unclear.OBJECTIVE: To investigate whether hUC-MSCs have an influence on the apoptosis of K562 cells induced by imatinib and to reveal the possible underlying mechanism.METHODS: K562 cells were cultured with hUC-MSCs or/and imatinib. Cellular apoptosis was measured with Annexin-V and PI staining by flow cytometry analysis. The protein expressions of Bax, Bcl-2, caspase-3, caspase-9 and cleaved-PARP in K562 cells were detected by western blot assay. Pan-caspase inhibitor Z-VAD-FMK was used to block apoptosis in each group, and during this process the effect of caspase apoptosis signaling pathway was detected.RESULTS AND CONCLUSION: The apoptosis of K562 cells was enhanced, when imatinib was combined with hUC-MSCs. Western blot analysis showed that the expression of pro-apoptotic protein Bax was enhenced and the expression of anti-apoptotic protein Bcl-2 was suppressed. Furthermore, the cleaved forms of caspase-9, caspase-3 and PARP in K562 cell were higher in the hUC-MSCs+imatinib group than in the imatinib group. The apoptosis of K562 cells induced by the hUC-MSCs combined with imatinib was significantly inhibited by Z-VAD-FMK. In conclusion, these findings indicate that hUC-MSCs can enhance imatinib-induced apoptosis of K562 cells by activating caspase apoptosis signaling pathway.

OBJECTIVETo explore the effects of principal-subordinate acupoints combination on improving myocardial ischemia, and the gene regulatory pathways for the protection of myocardial ischemia.

METHODSAccording to the random number table method, 70 SPF Wistar male rats were divided into a normal group, a model group, a LY294002 group, an insulin-like growth factors-1(IGF-1) group, a Neiguan group, an acupoint combination group and an acupoint combination + LY294002 group, 10 rats in each one. Rats in the normal group were injected with 0.9% NaCl solution, while rats in the remaining groups were treated with abdominal subcutaneous injection of isoroterenol hydrochloride to establish the rat model of myocardial ischemia. Rats in the LY294002 group and IGF-1 group were treated with injection of LY294002 solution and IGF-1 solution for 14 days. Rats in the Neiguan group were treated with electroacupuncture (EA) at "Neiguan" (PC 6) by using Han-200 EA apparatus for 10 min per treatment. Rats in the acupoint combination group were treated with EA at "Neiguan" (PC 6), "Zusanli" (ST 36) and "Guanyuan" (CV 4) by using Han-200 EA apparatus for 10 min per treatment. Rats in the acupoint combination + LY294002 group were treated with LY294002 solution for 14 days, and EA at "Neiguan" (PC 6), "Zusanli" (ST 36) and "Guanyuan" (CV 4) was given before model establishment, once a day for 21 days. EA pretreatment was given before model establishment in all acupuncture groups. The heart rate (HR) and ST segment voltage were detected before and after treatment; the myocardial pathological morphology was observed by HE staining; the expressions of P13K mRNA and Akt mRNA were tested.

RESULTSAfter modeling, HR and ST segment voltage in all intervention groups were higher than those in the normal group (all P < 0.01); after the intervention, the HR and the ST segment voltage in the acupoint combination group, IGF-1 group and IGF-1 group were improved (P < 0.01, P < 0.05), which was more significant in the acupoint combination group and Neiguan group (all P < 0.01). As for the myocardial pathological morphology, obvious myocardial ischemia was observed in the model group, and that in the LY294002 group was the most serious, and that in the acupoint combination+ LY294002 group was moderate. After intervention, the myocardial pathological damage in the IGF-1 group, Neiguan group and acupoint combination group was significant improved, which was more significant in the IGF-1 group and acupoint combination group. As for the expression of PI3K mRNA and Akt mRNA, compared with normal group, the expression of PI3K mRNA was increased in the remaining groups after modeling (P < 0.01, P < 0.05), which was more significant in the IGF-1 group and acupoint combination group (all P < 0. 01). The expression of Akt mRNA in the LY294002 group and acupoint combination + LY294002 group was reduced (P < 0. 01, P < 0.05), while that in the remaining groups was increased (P < 0.01, P < 0.05), which was more significant in the IGF-1 group and acupoint combination group (all P < 0.01).

CONCLUSIONThe principal-subordinate acupoints combination could improve heart rate and ST segment voltage in rats with chronic myocardial ischemia, reduce myocardial pathological damage, which is superior to single selection of "Neiguan" (PC 6). The PI3K/Akt signaling pathway may be involved in the regulation mechanism of principal-subordinate acupoints combination for the protection of chronic myocardial ischemia.

Acupuncture Points ; Acupuncture Therapy ; Animals ; Chronic Disease ; therapy ; Electroacupuncture ; Electrocardiography ; Heart Rate ; Humans ; Insulin-Like Growth Factor I ; metabolism ; Male ; Myocardial Ischemia ; enzymology ; pathology ; physiopathology ; therapy ; Myocardium ; pathology ; Phosphatidylinositol 3-Kinases ; genetics ; metabolism ; Proto-Oncogene Proteins c-akt ; genetics ; metabolism ; Rats ; Rats, Wistar

OBJECTIVETo explore the role of SCF/c-Kit signaling in the invasion of bladder cancer T24 cells.

METHODSWestern blotting was used to detect the expression of c-Kit and PI3K pathway activation stimulated by stem cell factor (SCF) in T24 cells. The invasiveness of T24 cells before and after SCF stimulation and Wortmannin (aspecific PI3K inhibitor) treatment was evaluated using Transwell invasion assay (direct and indirect counting methods).

RESULTST24 cells expressed c-Kit protein and showed obvious Akt phosphorylation after stimulation with SCF (1 ng/ml) for 24 h. Compared to the control group, SCF stimulation (1 ng/ml) caused a greater number of T24 cells to migrate through the polycarbonate film (P<0.01), and this effect was blocked by the application of Wortmannin before the stimulation.

CONCLUSIONSCF/c-Kit signaling promotes the invasiveness of T24 cells, and this effect is mediated by the PI3K pathway.

Carcinoma, Transitional Cell ; metabolism ; pathology ; Cell Line, Tumor ; Humans ; Neoplasm Invasiveness ; Proto-Oncogene Proteins c-kit ; metabolism ; Signal Transduction ; Stem Cell Factor ; metabolism ; Urinary Bladder Neoplasms ; metabolism ; pathology

Objective To explore the role of SCF/c-Kit signaling in the invasion of bladder cancer T24 cells. Methods Western blotting was used to detect the expression of c-Kit and PI3K pathway activation stimulated by stem cell factor (SCF) in T24 cells. The invasiveness of T24 cells before and after SCF stimulation and Wortmannin (aspecific PI3K inhibitor) treatment was evaluated using Transwell invasion assay (direct and indirect counting methods). Results T24 cells expressed c-Kit protein and showed obvious Akt phosphorylation after stimulation with SCF (1 ng/ml) for 24 h. Compared to the control group, SCF stimulation (1 ng/ml) caused a greater number of T24 cells to migrate through the polycarbonate film (P<0.01), and this effect was blocked by the application of Wortmannin before the stimulation. Conclusion SCF/c-Kit signaling promotes the invasiveness of T24 cells, and this effect is mediated by the PI3K pathway.

Objective To explore the role of SCF/c-Kit signaling in the invasion of bladder cancer T24 cells. Methods Western blotting was used to detect the expression of c-Kit and PI3K pathway activation stimulated by stem cell factor (SCF) in T24 cells. The invasiveness of T24 cells before and after SCF stimulation and Wortmannin (aspecific PI3K inhibitor) treatment was evaluated using Transwell invasion assay (direct and indirect counting methods). Results T24 cells expressed c-Kit protein and showed obvious Akt phosphorylation after stimulation with SCF (1 ng/ml) for 24 h. Compared to the control group, SCF stimulation (1 ng/ml) caused a greater number of T24 cells to migrate through the polycarbonate film (P<0.01), and this effect was blocked by the application of Wortmannin before the stimulation. Conclusion SCF/c-Kit signaling promotes the invasiveness of T24 cells, and this effect is mediated by the PI3K pathway.

Objective    To systematically review the impact of chronic kidney disease (CKD) at different stages on prognosis of transcatheter aortic valve replacement (TAVR). Methods    Databases including PubMed, the Cochrane Library, EMbase, Web of Science, CNKI, Wanfang and the Chinese Biomedical Literature Database (CBM) were searched by computer to collect cohort studies on impact of different stages of CKD on prognosis of TAVR from inception to July 2020. Two reviewers independently screened literature, extracted data and assessed risk of bias of included studies, and then, meta-analysis was performed by using Stata 15.0 software. Risk of study bias was assessed using the Newcastle-Ottawa Scale (NOS). Results    A total of 17 cohort studies were included with NOS score≥6 points. The results of meta-analysis indicated that: compared with the patients without CKD, all-cause mortality of CKD stage 3 patients at 30 day (RR=1.29, 95%CI 1.22-1.37, P<0.001) and 1 year (RR=1.24, 95%CI 1.19-1.28, P<0.001), all-cause mortality of CKD stage 4 patients at 30 day (RR=2.10, 95%CI 1.90-2.31, P<0.001) and 1 year (RR=1.89, 95%CI 1.62-2.19, P<0.001), and all-cause mortality of CKD stage 5 patients at 30 day (RR=2.22, 95%CI 1.62-2.19, P<0.001) and 1 year (RR=2.24, 95%CI 1.75-2.87, P<0.001) were significantly increased and were associated with the severity of CKD. The occurrence rates of 1-year cardiovascular mortality, postoperative acute kidney injury and bleeding events were all higher in patients with CKD. Conclusion    CKD at stages 3, 4 and 5 is associated with increased all-cause mortality after TAVR, and the higher the stage of CKD is, the higher the risk of all-cause mortality at 30-day and 1-year follow-up is. Due to limited quality and quantity of the included studies, more high-quality studies are required to verify above conclusions.