OBJECTIVE: To study the chemical fingerprint features and the quantitative analysis of 6 components of Fructus Trichosanthis and its steamed products. METHODS: The chemical fingerprints were established by HPLC with 10 batches of Fructus Trichosanthis and their steamed products, and the contents of 5-hydroxymethyl furfural, vanillic acid, rutin, isoquercitrin, quercetin and luteolin in Fructus Trichosanthis and their steamed products were quantitatively analyzed. The similarity of Fructus Trichosanthis and its steamed products was calculated by the correlation coefficient method and the included angle cosine method, and the contents of 6 components were tested by means of calibration curve. RESULTS: The 10 batch of Fructus Trichosanthis similarity was from 0.82 to 0.86, and the 10 batch of Fructus Trichosanthis steamed products similarity was from 0.97 to 0.98, there was significant difference between the two (P < 0.01). Compared with Fructus Trichosanthis, the contents of 5-hydroxymethyl furfural, vanillic acid, quercetin and luteolin content in steamed products showed significant changes (P < 0.05 or P < 0.01). CONCLUSION: Before and after Fructus Trichosanthis steamed, the change of fingerprint feature and changes of the component content provide experimental basis for further studies of Fructus Trichosanthis and its steamed products.

OBJECTIVE: To study the chemical constituents of different solvent extracts from Fructus Trichosanthis, and to study their antioxidant activity and protection of myocardial ischemia reperfusion injury in rats. METHODS: The chemical constituents in different solvent extracts were determined by UV-Vis spectrophotometry. Myocardial ischemia reperfusion model was made by ligation of left coronary artery in rats. Rats were randomly divided into 6 groups, with 6 rats in each group. The sham operation group and the model groups were respectively given normal saline, Fructus Trichosanthis (equivalent to crude drug 22.5 g·kg-1), compound Danshen dripping pill group(0.085 g·kg-1). Medicines were given once a day for 7 d. After the last drug 1 h, left coronary artery was ligated for 30 min and then reperfusion was established for 120 min by removing the ligation. During this time, ECG was recorded. At the end, animals were euthanized. Blood was collected to evaluate the contents of CK-MB, MYO-MB, cTnT. The heart was removed and fixed to observe the changes of myocardial tissue by optical microscope. RESULTS: Compared with the water extract group and the alcohol extract group of Fructus Trichosanthis, the total amino acids content of Fructus Trichosanthis dichloromethane extract was not detected, but the content of total flavonoids is higher (P < 0.01). Compared with the water extraction liquid of Fructus Trichosanthis, the antioxidant properties on DPPH radical of Fructus Trichosanthis dichloromethane extract group is lower (P < 0.01), and the antioxidant activity on-OH free radical of the alcohol extract of Fructus Trichosanthis group is lower (P < 0.01). Compared with model group, the elevation of ST segment of electrocardiogram was significantly suppressed in each group during reperfusion (P < 0.01 or P < 0.05). The plasma CK-MB, cTnT and MYO-MB in water extract group were significant lowered (P < 0.05). CONCLUSION: The extraction of Fructus Trichosanthis is able to decrease the production of oxidants. The water extract of Fructus Trichosanthis could ameliorate myocardial ischemia reperfusion injury.

OBJECTIVE: To study the effects of rice wine on the metabolism of 3, 29-dibenzoyl karounitriol as the main active component in Gualou-Xiebai in human intestinal flora, exploring the scientific connotation of Gualou-XiebaiBaijiu Decoction from the angle of metabolism. METHODS: The metabolic rate of 3, 29-dibenzoyl karounitriol in Gualou-Xiebai rice wine solution and aqueous solution at different time points were compared,respectively,by intestinal flora experiments. RESULTS: Compared to Gualou-Xiebaiaqueous solution group, the metabolic rate of 3, 29-dibenzoyl karounitriol in Gualou-Xiebai rice wine solution group decreased significantly (P<0.05). CONCLUSION: Rice wine as cites drug can inhibit the metabolism of 3, 29-dibenzoyl karounitriol in Gualou-Xiebai.

Objective To study the similarity of chemical fingerprints of different proportion of Gualoupi-Gualouzi and Fructus Trichosanthis, so as to provide the theoretical basis for the reasonable proportion of Gualoupi-Gualouzi. Methods The HPLC fingerprints and the common patterns of 6 the proportion of the 6 kinds of compatibility (30∶70, 35∶65, 40∶60, 45∶55, 50∶50 and 55∶ 45) of Gualoupi-Gualouzi and Fructus Trichosanthis were established, and the similarity of chemical fingerprints of different compatibility proportion of Gualoupi-Gualouzi and Fructus Trichosanthis between their common patterns were compared. Results Compared with the common pattern of Fructus Trichosanthis, the similarities of the common patterns of chemical fingerprints of the above 6 kinds of compatibility proportion of Gualoupi-Gualouzi were 0.8808±0.0407, 0.8930±0.0236, 0.8995±0.0195, 0.9033±0.0190, 0.9363± 0.0082 and 0.8904±0.0144, respectively. Among them, Gualoupi-Gualouzi ratio of 50∶50 had the highest similarity, and compared with other groups, the differences were statistically significant (P<0.05 or 0.01). Conclusion The optimum proportion of Gualoupi- Gualouzi can be determined by the fingerprint technology, which provides the theoretical basis for the reasonable proportion of Gualou⁃pi-Gualouzi.

Objective To verify the protective effects of Gualou Xiebai dropping pills(GX) on myocardial ischemia-reperfusion injury(MIR.I) in rats. Methods After screening the qualified SPF rats were randomly divided into four groups, with 12 in each group. Sham-operated group and model group rats were respectively treated with normal saline, and rats in GX group and compound Danshen dropping pills group were given the corresponding dropping pills equivalent to 22.5 g/kg and 85.05 mg/kg of the crude herb, respectively. All groups were administered once a day for 7 successive days. One hour after the last administration, the MIRI models were produced by occluding the left coronary artery for 30 min and releasing the occlusion for 120 min. The changes in ST segment were observed, and the contents of creatine kinase-MB (CK-MB), cardiac troponin-T (CTNT) and myoglobin (MYO) in blood plasma were measured. The pathological changes in myocardial tissues were observed by optics and electric-microscope and the percentage of myocardial infarction was measured by detecting the content of Evan blue in myocardium. Results Compared to normal saline in the model group, GX had antagonism to ECG S-T segments elevation in rats with myocardial ischemia, while the contents of CK-MB, MYO, and CTNT in blood plasma declined significantly (P<0.05 or P<0.01); the disorder condition of myocardial cell fiber arrangement was improved, and edema between myocardial tissue and cells was relieved; the inflammatory cell infiltration and myocardial necrosis in cardiac allograft were significantly alleviated, and the percentage of myocardial infarction was decreased significantly (P< 0.01). Conclusion GX may play an important protective role against the MIRI in rats.

Objective: To explore whether the basic principle of Fisher component analysis(FCA) can be used in the category analysis of Fructus Trichosanthis fruit and its processed products. Methods: Their fingerprints were established by using HPLC-PDAD, and the standard fingerprints of them were obtained by digitizing with quercetin as the internal reference peak. Then, their chemical fingerprint information was extracted by FCA and classified by quality model, and the classification results were compared with those classified by principal component analysis and standard atlas analysis. Results: Fructus Trichosanthis and its processed products can be accurately divided into two categories by FCA. Conclusion: FCA can extract the implied chemical information in fingerprints, and analyze them accurately.

Objective To study the in situ intestinal absorption behaviors of 3, 29- dibenzoyl- karounitriol (DK) in Gualou- Xiebai (GX) extract in rats. Methods A rat in situ single-pass perfusion model was used and the concentrations of the perfusate were determined by HPLC-PDAD to investigate the intestinal absorption site and mechanism. Results The main absorption site of DK in GX extract was jejunum, ileum and colon, and the absorption had no significant difference in the three different segments of rat intestine (P>0.05), but was significantly higher than that in duodenum (P<0.05). The Ka and Papp of DK had no significant difference in different concentrations of GX extract (P>0.05). Conclusion DK in GX extract could be absorbed in whole intestinal segment, with the best intestinal absorption site of jejunum, ileum and colon. Its absorbing mechanism may be related to passive diffusion.

Objective To explore whether evolving window orthogonal projection analytical method can be used in the chromatographic peaks matching between traditional Chinese medicine preparation and its medicinal ingredients in prescription. Methods The fingerprints of Gualou Xiebai Baijiu Decoction and the herbs (Gualou and Xiebai) in its prescription were established by using high performance liquid chromatography-photodiode array detector (HPLC-PDAD), and the HPLC peaks in fingerprints of Gualou Xiebai Baijiu Decoction and the herbs in its prescription were matched according to the principle of evolving window orthogonal projection analytical method. Results Evolving window orthogonal projection analytical method could match the chromatogram peaks well between different fingerprints of Gualou Xiebai Baijiu Decoction and the herbs in its prescription. There were 20 common peaks between the ethyl acetate extraction of Gualou Xiebai Baijiu Decoction and Gualou, and 9 common peaks between the n-butanol extraction of Gualou Xiebai Baijiu Decoction and Xiebai. Conclusion Evolving window orthogonal projection analytical method can be used in the chromatographic peaks matching between traditional Chinese medicine preparation and its medicinal ingredients in prescription.

Cell Transformation, Neoplastic ; drug effects ; genetics ; Dimethyl Sulfoxide ; pharmacology ; Electroporation ; methods ; HL-60 Cells ; Humans ; Membrane Proteins ; genetics ; Neoplasm Proteins ; genetics ; RNA Interference ; RNA, Small Interfering ; genetics ; Stromal Interaction Molecule 1 ; Transfection

To investigate the role of ion channels in the coupling responses of neutrophils to extracellular stimulus, it is necessary to study the membrane ion channel activities using patch-clamp technique. However, little has been known about the ion channel activities in neutrophils due to the difficulties in forming giga-seal with pipettes because of small diameter of neutrophils and the easily developed polarization. Some studies indicated that favorable results could be achieved through pretreatment at low temperature before electrophysiological recordings. But it remains unclear whether the pretreatment affects the membrane current and why the seal rate increases after low temperature pretreatment. The purpose of this study was to investigate the effects of 4 degrees C pretreatment on the membrane current and cell polarity in human neutrophils. In the experiments, human neutrophils were isolated from fresh peripheral blood of healthy volunteers and divided into two groups (room temperature group and 4 degrees C pretreatment group). Voltage-dependent K(+) (Kv) currents were recorded in whole-cell voltage-clamp mode and large-conductance Ca(2+)-activated K(+) (BK(Ca)) currents were recorded using inside-out patches. The results showed that 4 degrees C pretreatment significantly inhibited cell polarity (P<0.05), and it took more time for neutrophils to form a polarity-cycle [(534+/-32) s, n=20] compared with those at room temperature [(257+/-24) s, n=20]. Meanwhile, seal rate significantly increased in 4 degrees C pretreatment group (64%) compared with that in the room temperature group (27.5%). The seal rate and cell polarity rate during 0 approximately 1 min after 4 degrees C pretreatment were significantly different from those at room temperature, while no significant difference was found during 9 approximately 10 min between the two groups. Our results suggest that 4 degrees C pretreatment can inhibit cell polarity and increase seal rate, but has no effects on membrane currents. It is also suggested that 0 approximately 1 min after 4 degrees C pretreatment is a more suitable time for electrophysiological recording in neutrophils.
Cell Polarity ; Cold Temperature ; Humans ; Large-Conductance Calcium-Activated Potassium Channels ; physiology ; Membrane Potentials ; Neutrophils ; physiology ; Potassium Channels, Voltage-Gated ; physiology