Myricetin inhibits interferon-γ-induced programmed death ligand-1 and indoleamine 2, 3-dioxygenase 1 expression in lung cancer cells / 中国药理学与毒理学杂志

OBJECTIVE Programmed death ligand-1 (PD-L1) and indoleamine 2, 3-dioxygenase 1 (IDO1) are immune checkpoints which can be induced by interferon-γ(IFN-γ) in the tumor microenvironment, leading to immune escape of tumors. Myricetin (MY) is a flavonoid distributed in many edible and medicinal plants. The aim of this study is to clarify the effect and the mechanism of MY on inhibiting IFN-γ-induced PD-L1 and IDO1 in lung cancer cells. METHODS Expressions of PD-L1 and major histocompatibility complex-I (MHC-I) were evaluated by flow cytometry and Western blotting, and the expression of IDO1 was measured by Western blotting. qRT-PCR was used to detect their mRNA levels. The function of T cells was evaluated using a co-culture system consist of lung cancer cells and the Jurkat-PD-1 T cell line that overexpressing PD-1. Molecular docking analysis, Western blotting and immunofluorescence were used for mechanism study. RESULTS MY potently inhibited IFN-γ-induced PD-L1 and IDO1 expression in human lung cancer cells, while didn't show obvious effect on the expression of MHC-I. In addition, MY restored the survival, proliferation, CD69 expression and interleukin-2 (IL-2) secretion of Jurkat-PD-1 T cells suppressed by IFN-γ-treated lung cancer cells in the co-culture system. Mechanistically, IFN-γ up-regulated PD-L1 and IDO1 at the transcriptional level through the JAK-STAT-IRF1 axis, which was targeted and inhibited by MY. CONCLUSION Our research revealed a new insight into the anti-tumor effects of MY which inhibited IFN-γ-induced PD-L1 and IDO1 expression, supporting the potential of MY in anti-tumor immunotherapy.

Gastric dynamics analysis of TEAS combined SNP induced general anesthesia when controlled hypotension dropped to 60% of the MAP baseline / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To observe the effect of gastric dynamics by transcutaneous electrical acupoint stimulation (TEAS) combined general anesthesia when controlled hypotension dropped to 60% of the mean arterial prenssure (MAP) baseline, and to provide experimental evidence for organ protection in clinical controlled hypotension.</p><p><b>METHODS</b>Eighteen male beagles were randomly divided into three groups, the general anesthesia group (blank), the general anesthesia induced controlled hypotension group (control), and the general anesthesia combined TEAS induced controlled hypotension group (experiment), 6 in each group. Controlled hypotension was performed in the latter two groups with isoflurane inhalation and intravenous injection of sodium nitroprusside (SNP). The mean arterial pressure (MAP) was lowered to 60% of the MAP baseline and kept for 60 min. Controlled hypotension was not performed in Beagles of the control group. For Beagles in the experiment group, TEAS [2/100 Hz, (4 ± 1) mA] was applied to bilateral Hegu (LI4), Quchi (LI11), Zusanli (ST36), and Sanyinjiao (SP6) from stable physiological conditions to the end of maintaining stages. Changes of EGG frequencies and EGG amplitudes were monitored. Serum levels of gastrin (GAS) and motilin (MTL) were also detected at corresponding time points during and after experiment.</p><p><b>RESULTS</b>As for the pressure control effect of TEAS combined general anesthesia in the controlled hypotension, during the process of controlled hypotension (T1-T4), MAP levels of two controlled pressure groups remained relatively stable, and were kept at 60% of the MAP baseline. When the blood pressure dropped to the target low MAP and maintained at 60 min (T1-T4), EGG amplitudes of Beagles in all the three groups showed decreasing tendency. But it was more obviously lower than its basic level in the control group (P <0.05), while it was not obviously decreased in the experiment group (P < 0.05). EGG frequencies of Beagles in all the three groups showed no obvious change during this stage. By the end of the MAP rising stage (T8), the EGG amplitude of the experimental group was significantly higher than that of the control group and the blank group (P < 0.05), while it didn' t show any obvious increase in the control group. During this period, EGG frequencies of the two controlled hypotension groups decreased more than those of the blank group. Two h after rising blood pressure (at T9), EGG amplitudes and frequencies in the two controlled hypotension groups basically restored to their respective baselines and levels of the blank group at T9. At 2 h (T9) after controlled hypotension, serum levels of GAS and MTL were lower than those of basic levels in the two controlled hypotension groups (P <0.05). However, serum levels of GAS and MTL had an increasing trend in the two controlled hypotension groups at 24-72 h (T10-T12). Besides, the increasing speed and amplitude was better in experiment group than in the control group at T10-T12. However, there was no statistical difference between the two groups (P > 0.05). At 72 h (T12) serum levels of GAS and MTL had basically restored to their basic levels in the two controlled hypotension groups and that of the blank control group.</p><p><b>CONCLUSION</b>EGG amplitudes could be effectively improved in TEAS combined general anesthesia for controlled hypotension at 60% of the MAP baseline, the recovery of the serum GAS level accelerated, gastric power improved and stomach protected.</p>

Screening bioactive compounds inhibiting influenza virus from isatidis radix by ultrafiltration mass spectrometry / 中国中药杂志

In vitro neuraminidase inhibition assays and ultrafiltration liquid chromatography with diodearray detector coupled to time of flight mass spectrometer (UPLC-DAD-TOF-MS) were combined to screen bioactive compounds inhibiting neuraminidase from Isatidis Radix. By comparing the compounds from Isatidis Radix before and after ultrafiltration, we found that arginine, goitrin and adenosinea can bind with neuraminidase, and the binding degree of the three compounds were (36.23 +/- 1.12)%, (32.54 +/- 1.02)% and (9.38 +/- 0.47)%, respectively. The IC50 of arginine and goitrin were (1.16 +/- 0.02), (1.20 +/- 0.02) g x L(-1), respectively. While the IC50 of adenosinea was higher than 500 g x L(-1). The results showed that arginine and goitrin might be the main compounds with antiviral activity of Isatidis Radix. This study may provide a useful method for the screening of bioactive compounds and quality control of Isatidis Radix.

Dynamic monitoring of the cytotoxic effects of protoberberine alkaloids from Rhizoma Coptidis on HepG2 cells using the xCELLigence system / 中国天然药物

AIM@#To investigate the cytotoxic effects of the six protoberberine alkaloids (PAs) from Rhizoma Coptidis on HepG2 cells.@*METHOD@#A systematic screening was conducted to investigate the dynamic response of HepG2 cells to the PAs using the impedance-based xCELLigence system. Cisplatin was selected as the positive control. The real time, concentration-response curves and the 50% inhibitory concentrations (IC50) were acquired to evaluate the anticancer activity of the PAs.@*RESULTS@#All of the six PAs inhibited cell growth and induce death in HepG2 cells in a time- and concentration-dependent manner. The IC50 values of cisplatin, berberine, columbamine, coptisine, epiberberine, jatrorrhizine, and palmatine were 5.13, 42.33, 226.54, 36.90, 302.72, 383.54, and 456.96 μg·mL(-1), respectively. The results obtained using the xCELLigence system corresponded well with those of the conventional methods.@*CONCLUSION@#The xCELLigence system is a reliable and efficient tool for real-time screening of the cytotoxic effect of compounds in cell-based in vitro assays. Coptisine and berberine, with methylenedioxy group at C2 and C3 on the phenyl ring showed stronger effect.than the other four PAs. However, compared with cisplatin, the six PAs didn't show obvious cytotoxic effect on HepG2 cells. These results provided some useful data for the evaluation of the anticancer compounds, and the clinical application of traditional Chinese medicine.

A novel method for testing sterility of injections based on biothermodynamics / 药学学报

This study aims at trying to establish a novel method of sterility test for injections based on biothermodynamics, in order to overcome the deficiencies of routine sterility tests such as long detecting cycle, low sensitivity and prone to misjudgments. A biothermodynamics method was adopted to rapidly detect the microorganism contamination of injections by monitoring the heat metabolism during the growth of microbe. The growth rate equal to or greater than zero and the heat power difference of P(i) and P(0) with three folds higher than the noise of baseline were chosen as indexes to study the heat change rule of microbe. In this way, the effectiveness of the new method to detect strains required by conventional sterility test or in injection samples was also investigated. Results showed that the Gram-positive bacteria, Gram-negative bacteria and fungi demanded by sterility testing methodology could be detected by biothermodynamics method within 10 hours, with the sensitivity lower than 100 CFU x mL(-1). Meanwhile, this method was successfully applied to the sterility test of Compound Yinchen injection (FFYC), Shuanghuanglian powder injection (SHL) and Compound Triamcinolone injection (TAND) which were sterilized with different degrees. Therefore, the biothermodynamics method, with advantages of fast detection and high sensitivity, could be a complementary solution for conventional sterility tests.

Microcalorimetric investigation on the interaction of six alkaloids from rhizoma coptidis / 药学学报

How to identify active constituents of traditional Chinese medicines (TCMs) and study their interactions are key problems in the development of TCMs. The inhibitory effect of six alkaloids from Rhizoma Coptidis (RC) on Shigella dysenteriae (S. dysenteria) growth had been investigated by microcalorimetry in this study. Main active constituents of RC were confirmed by comparing their contributions to the bacteriostatic effect, and the interactions among active constituents were further researched. According to the result, in 0.8 mg-mL-1 extract of RC, the contributions of six active alkaloids including berberine, coptisine, epiberberine, palmatine and the combination of jatrorrhizine and columbamine were 52.83%, 36.31%, 2.49%, 4.27% and 3.21%, respectively. Therefore, berberine and coptisine were the main active constituents of RC that inhibited the growth of S. dysenteria. The study of interactions among the six alkaloids indicated that, 1 there were some contstituents antagonizing the inhibitory effect of RC, 2 there was a synergy effect between berberine and coptisine, 3 there were additive effects between other four alkaloids and the main active constituents. These results may provide some useful references for the establishment of the quality standard for RC and the development of multi-component TCMs.

Effects of transcutaneous electrical acupoint stimulation combined general anesthesia in controlled hypotension at different levels on the liver blood flow / 中国中西医结合杂志

<p><b>UNLABELLED</b>OBJECTIVE To observe the changes of the liver blood flow in controlled hypotension by transcutaneous electrical acupoint stimulation (TEAS) combined general anesthesia, thus clarifying the mechanism of liver protection effect in acupuncture anesthesia combined with drugs.</p><p><b>METHODS</b>Forty-two male beagles were randomly divided into seven groups, i.e. , the general anesthesia group, the 50% control group, the 50% experiment group, the 40% control group, the 40% experiment group, the 30% control group, and the 30% experiment group, 6 in each group. Beagles in the latter six groups were administered with isoflurane inhalation and intravenous dripping of sodium nitroprusside (SNP) for controlled hypotension. The mean arterial pressure (MAP) was lowered to 50%, 40%, and 30% basic MAP and lasted for 60 min. Beagles in the general anesthesia group was not treated with controlled hypotension. In the experiment groups, TEAS was applied to bilateral Hegu (LI4), Zusanli (ST36), Sanyinjiao (SP6), and Quchi (LI11) at 2/100 Hz with the stimulation strength of 4 +/- 1 mA. The TEAS started from the stability of physiological conditions to 60 min after maintaining the target MAP. The changes of blood flow of the liver tissue surface at corresponding time points were monitored by laser Doppler blood flow meter.</p><p><b>RESULTS</b>Between the beginning of hypotension and the maintaining stage of target low blood pressure, the liver blood flow of the 50% control group was significantly lower than the level of the general anesthesia group and the basic level at corresponding time points (P < 0.05). It was significantly reduced in the 50% experiment group only at 30-60 min of maintenance. Besides, in the early period of maintenance (10-30 min), it was significantly higher in the 50% experiment group than in the 50% control group at the same time points (P < 0.05). In this stage, there was no obvious increase in the liver blood flow in the 40% and 30% experiment groups. In the recovery phase of blood pressure (20-30 min), the liver blood flow of the 40% experiment group had been restored to the level of the 40% control group and the basic level, while it had not been restored in the general anesthesia group. In this stage, the similar changing tendency of the liver blood flow occurred in the 50% and 30% experiment groups and the 50% and 30% control groups.</p><p><b>CONCLUSIONS</b>Line to a high level of controlled hypotension (50%), TEAS liver protective effect was obviously embodied in the early step-down phase and the maintenance phase. Line-induced hypotension to a lower level (40%), TEAS liver protective effect was obviously embodied in the recovery phase.</p>

Effects of the renal blood flow at different levels by transcutaneous electrical acupoint stimulation combined general anesthesia induced controlled hypotension / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To observe the intervention of transcutaneous electrical acupoint stimulation (TEAS) on the renal blood flow at different levels of mean arterial pressure (MAP) in controlled hypotension.</p><p><b>METHODS</b>Forty-two male beagle dogs were randomly divided into seven groups, i. e., the general anesthesia group, the 50% controlled group, the 40% controlled group, the 30% controlled group, the 50% experimental group, the 40% experimental group, and the 30% experimental group, 6 in each group. Beagles in the general anesthesia group were not treated with controlled hypotension, and the target MAP was achieved in those of the rest groups and maintained for 60 min. In the experimental groups, TEAS was applied to bilateral Hegu (LI4), Zusanli (ST36), Sanyinjiao (SP6), and Quchi (LI11) at 2/100 Hz with the stimulation strength of (4 +/- 1) mA starting from the stability of their physiological conditions to 60 min of maintaining the target MAP level. The changes of the renal blood flow were monitored at different time points using laser Doppler.</p><p><b>RESULTS</b>From starting pressure control to the target MAP level, the renal blood flow was significantly lower in the 30% controlled group than in the general anesthesia group and the basic level of the same group (P < 0.05), while there was no obvious change in the 30% experimental group. In maintaining the blood pressure, the renal blood flow was significantly lower in the 50% controlled group, the 40% controlled group, the 30% controlled group, and the 30% experimental group than in the general anesthesia group (P < 0.05), while there was no obvious change in the 50% experimental group or the 40% experimental group. By the end of blood pressure recovery, the renal blood flow restored to the basic level in the 50% controlled group, the 50% experimental group, and the 40% experimental group (P > 0.05), while it was not restored to the basic level in the 40% controlled group, the 30% controlled group, and the 30% experimental group (P < 0.05).</p><p><b>CONCLUSION</b>TEAS combined general anesthesia in controlled hypotension could effectively improve the renal blood flow, thus protecting the kidney.</p>

Effects of transcutaneous electrical acupoint stimulation combined with general anesthesia for controlled hypotension on heart function and oxidation resistance of myocardium / 中国针灸

<p><b>OBJECTIVE</b>To investigate the intervention effect of transcutaneous electrical acupoint sitmulation (TEAS) on heart under limiting level of controlled hypotension.</p><p><b>METHODS</b>Eighteen male Beagles were randomly divided into a general anesthesia group, a control group and an experimental group, 6 dogs in each group. All animals were applied with general anesthesia, and the animals of the control group and the experimental group were administered with combination of isoflurane and sodium nitroprusside (SNP) for controlled hypotension to 30% mean arterial pressure (30% MAP) of base line for 60 min. In the experimental group, TEAS (2 Hz/100 Hz, 6-8 mA) was applied to "Hegu" (LI4), "Zusanli" (ST 36), "Sanyinjiao" (SP 6) and "Quchi" (LI 11) from the beginning of physiological conditions stability to the end of maintained low MAP for 60 min. Creatine kinase isoenzymes MB (CK-MB) and lactate dehydrogenase (LDH) activity were detected at the base line and 2 h, 24 h, 48 h, 72 h after operation. Total superoxide dismutase (T-SOD) and malondialdehyde (MDA) in myocardium were observed at 72 h after operation.</p><p><b>RESULTS</b>The peak of LDH arrived at 24 h after operation and then restored in 3 groups. Compared with basal level, the activity of LDH in controlled group at 72 h after operation was significantly increased (P < 0.05), and in the other two groups no obvious change (P > 0.05). The activity of CK-MB arrived to peak from 2 h to 24 h after operation, and then came back gradually in 3 groups. Compared with postoperative 72 h of the same group, the control group at 48 h significantly increased (P < 0.05), while the other two groups had no statistically significant difference (P > 0.05). The content of MDA at 72 h after operation in controlled group was higher significantly than that in the general anesthesia group and the experimental group (both P < 0.05).</p><p><b>CONCLUSION</b>TEAS combined with anesthesia for controlled hypotension can elevate myocardial effect of anti-oxide free radical, reduce the postoperative release of LDH and CK-MB, so as to produce myocardial protection.</p>

Cardiac protective effects of transcutaneous electrical acupoint stimulation combined with general anesthesia for controlled hypotension / 中国针灸

<p><b>OBJECTIVE</b>To explore the protective effect and the mechanism of controlled hypotension induced by transcutaneous electrical acupoint stimulation (TEAS) combined with general anesthesia.</p><p><b>METHODS</b>Sixteen male Beagles were randomly divided into a group of controlled hypotension induced by simple general anesthesia (control group) and a group of controlled hypotension induced by TEAS combined with general anesthesia (observation group). All the animals were administered with combination of Isoflurane and Sodium Nitroprusside (SNP) for controlled hypotension without TEAS until the arterial pressure was lowered to 30% basic mean arterial pressure (MAP) for 60 min. In the observation group, TEAS (2 Hz/100 Hz, 3-5 mA) was applied to "Hegu" (LI 4) "Zusanli" (ST 36), "Sanyinjiao" (SP 6) and "Quchi" (LI 11) from the beginning of physiological conditions stability to the end of maintained low MAP for 60 min, but there was no TEAS in control group. The changes of MAP, the left intraventricular pressure (LIVP), T wave and ST-T segment of II lead electrocardiogram (ECG) were monitored with the physiological signal acquisition system, and myocardial apoptosis was detected by TUNEL method.</p><p><b>RESULTS</b>All the animals could maintain stable required low blood pressure. At one hour after cease of controlled hypotension, MAP of (109.56 +/- 6.14) mmHg returned to the basic level in the observation group, while MAP of (84.91 +/- 6.36) mmHg was still lower than its basic MAP of (111.02 +/- 4.15) mmHg in the control group (P < 0.05), showing significant difference in MAP between the two groups (P < 0.05). -dp/dtmax of (3156.32 +/- 332.82) mmHg/s showed significant lower than its basic value of (4585.33 +/- 638.55) mmHg/s when blood pressure increased for 1 h in the control group (P < 0.05), but there was no difference in the observation group. When the objective low MAP maintaining for 60 min the ST segment was decreased significantly in the control group (P < 0.05), but there was no difference in the observation group. The numbers of positive apoptosis cardiocytes in the observation group were (24.67 +/- 2.45) cells/mm2, which were significantly fewer than (37.89 +/- 1.90) cells/mm2 in the control group (P < 0. 05).</p><p><b>CONCLUSIONS</b>TEAS combined with general anesthesia for controlled hypotension can significantly shorten restoration time of MAP, help to improve myocardial ischemia and the cardiac functional recovery and reduce myocardial apoptosis so as to produce myocardial protection.</p>

Freeze-drying of silymarin-loaded solid lipid nanoparticles (SM-SLN) / 中国中药杂志

<p><b>OBJECTIVE</b>To investigate lyophilization of SM-SLN.</p><p><b>METHOD</b>The parameters of lyophilization process was optimized. In addition, the protective effect of various types and concentrations of cryoprotectants were tested by shape, colour and disparity.</p><p><b>RESULT</b>The mixture of 2% lactose and 2% glucose could better prevent nanoparticles from aggregating, the optimal lyophilization process was followed: precooled at -45 degrees C for 10 hr; primary drying at -25 degrees C for 5 hr; secondary drying at 10 degrees C for 3 hr; finally drying at 30 degrees C for 6 hr.</p><p><b>CONCLUSION</b>Changes in particle size distribution during lyophilization could be minimized by optimizing the parameters of the lyophilization process and adding supporting agent.</p>