[Analysis of lignans and their metabolites derived from Schisandra chinensis and vinegar Schisandra chinensis in rats' plasma, bile, urine and faeces based on UHPLC-QTOF/MS].

UHPLC-QTOF/MS technique was used to study the differences of lignans and their metabolites derived from Schisandra chinensis and vinegar Schisandra chinensis in rat plasma, bile, urine and faeces by the data processing techniques such as the dynamic background subtract(DBS), mass defect filtering(MDF) and enhance peak list (EPL) in analysis. In order to enhance accuracy for Schisandra chinensis hepatoprotective effect, we established rat acute alcoholic liver injury model in this experiment, and studied the prototype components and metabolisms of Schisandra lignans in vivo under pathological condition. The main ingredients of alcohol extract are lignans, including deoxyschizandrin, schisandrin B, schizandrin C, schizandrol, schizandrol B,schisantherin, schisantherin B, schisanhenol, gomisin G, gomisin J. The metabolic transformation of lignans in rats was mainly induced by methylation, hydroxyl, oxidation, and so on. Finally, we identified 6 kinds of prototype components and their 20 potential metabolites in Schisandra chinensis group and vinegar Schisandra chinensis group.

[Effect of Astragali Radix with different sulfur fumigation technologies on immune function in mice].

To investigate the effect of Astragali Radix with different sulfur fumigation technologies on immune function in mice, and observe the effect of different Astragali Radix samples on carbon clearance in cyclophosphamide induced immunosuppressed mice, on immune organ weight in immunosuppressed mice and on delayed type hypersensitivity response (DTH) induced by 2,4 dinitrochlorobenzene. Carbon clearance index, phagocytic index, organ index and ear swelling rate were taken as the indexes. The results showed that, all of the Astragali Radix with different sulfur fumigation technologies markedly increased the carbon clearance index K, phagocytic index α, immune organ weight and improved the ability of DTH response in immunosuppressed mice. As compared with the model group, combined hot air-microwave group had the most significant difference, but when other groups were compared with and combined hot air-microwave group, only carbon clearance test had significant difference. From the perspective of pharmacodynamics, the effect of Astragali Radix with different sulfur fumigationon technologies on the immune function of mice was investigated, which provided a reference for the selection of appropriate alternative technology, and also provided guidance for clinical medication.

[Neural apoptosis and apoptosis-related genes in intracerebral hemorrhage patients].

OBJECTIVE: To observe the neural apoptosis and expression of apoptosis-related genes in brain in order to elucidate the regulation mechanism in the perihematomal region of intracerebral hemorrhage (ICH) patients. METHODS: Specimens of perihematomal region in human brain were obtained from 29 patients undergoing surgical evacuation of an intracerebral hematoma. Specimens of brain tissue were collected from the corpses of 6 persons within 3 hours after the accidental death. Neural apoptosis was evaluated by terminal deoxynucleotidyl transferase-mediated deoxyuridine 5' triphosphate nick end labeling (TUNEL) method and immunohistochemistry was used to detect the expression of Bcl-2, Bax, P53, and caspase-3 genes. RESULTS: The apoptosis rates of the ICH group was 4.10 +/- 0.28, significantly higher than that of the control group (0.57 +/- 0.43, P < 0.01). The expression rate of Bcl-2 the ICH group was 2.68 +/- 0.52, significantly higher than that of the control group (1.54 +/- 0.56, P < 0.01). The expression rate of Bax of the ICH group was 3.49 +/- 0.18, significantly higher than that of the control group (0.96 +/- 0.27, P < 0.01). The expression of P53 was 4.12 +/- 0.63, significantly higher than that of the control group (0.96 +/- 0.71, P < 0.01). The expression of caspase-3 of the ICH group was 3.50 +/- 0.25, significantly higher than that of the control group (0.74 +/- 0.73, P < 0.01). The expression levels of Bcl-2 and P53 were negatively correlated with the apoptosis rate (both P < 0.01), while the expression levels of Bax and caspase-3, and the Bax/Bcl-2 ratio were positively correlated with the apoptosis rate in perihematomal region of ICH patients (all P < 0.01). CONCLUSION: Apoptosis is involved in the delayed brain injury after ICH in human and is the main factor of delayed neural death. Some of the genes take part in the regulation of neural apoptosis: Bax and caspase-3 hasten the apoptosis while Bcl-2 and P53 restrain it.

[Effects of electroacupuncture on pain reactions, expression of spinal kappa-opioid receptor and contents of enkephalin and beta-endorphin in periaqueductal gray of midbrain in dysmenorrhea model rats].

OBJECTIVE: To observe effects of electroacupuncture (EA) on the expression of kappa-opioid receptor in the dorsal horn of the spinal cord and contents of enkephalin(ENK) and beta-endorphin (beta-EP) in the periaqueductal gray (PAG) of midbrain in dysmenorrheal rats, so as to reveal its underlying mechanism in relieving dysmenorrhea. METHODS: A total of 80 female SD rats were randomized into saline control (control), model, Sanyinjiao (SP 6), Xuanzhong (GB 39), non-acupoint groups (16 rats/group). Dysmenorrhea model was established by subcutaneous injection of estradiol benzoate (0.5 mg/rat on the 1st day and 10th day, 0.2 mg/rat from the 2nd day to the 9th day). One hour after the last injection, oxytocin (2 U/rat) was given intraperitoneally, for rats of the control group, the same dose of saline was given (i. p.). On the 10th day, EA (2 Hz/100 Hz, 0.1-0.3 mA) was applied to "Sanyinjiao" (SP 6), "Xuanzhong" (GB 39) and non-acupoint (the mid-point between the Stomach Meridian and Gallbladder Meridian, and in parallel with GB 39) for 20 min, respectively. Latency and number of writhing response, and writhing score (according to Schmauss's and Yaksh's method) were recorded. The expression of kappa-opioid receptor (kappa-OR) in T13, L1 , L2, L6 and S1 segments of spinal cord was determined by immunohistochemistry, and the contents of ENK and beta-EP in the midbrain PAG were assayed by ELISA. RESULTS: (1) Compared with the saline control group, the writhing latency of the model group was significantly shortened (P < 0.01), while the writhing times and writhing score of the model group were increased significantly (P < 0.01). Compared with the model group, the writhing latency of SP 6 group was significantly prolonged (P < 0.05), while the writhing scores and writhing times of the SP 6, GB 39 and the non-acupoint groups decreased significantly (P < 0.01). (2) In comparison with the control group, kappa-OR expression in the dorsal horn of L2 segment of spinal cord was upregulated significantly in the model group (P < 0.05). Compared to the model group, kappa-OR expression levels in the dorsal horns (DHs) of spinal T13, L1, L2, L6 and S1 segments in the SP 6 group were upregulated significantly (P < 0.01). ENK and beta-EP contents of PAG in the SP 6 and GB 39 groups were increased considerably (P < 0.01, P < 0.05). The effects of SP 6 were significantly superior to those of GB 39 in upregulating kappa-OR expression of spinal L1, L2 and L6 DHs and in upregulating beta-EP content of PAG; and superior to non-acupoint in upregulating kappa-OR expression of spinal T13, L1, L2, L6 and S1 DHs and in increasing both ENK and beta-EP contents of PAG (P < 0.01, PF < 0.05). No significant differences were found between the non-acupoint group and the model group in writhing latency, kappa-OR expression levels of spinal T13, L1, L2 and S1 DHs, and in ENK and beta-EP contents of PAG (P > 0.05). CONCLUSION: EA of SP 6 can significantly alleviate pain reactions in dysmenorrhea rats, which is closely associated with its functions in upregulating spinal kappa-OR expression and ENK and beta-EP contents in PAG. EA of SP 6, GB 39 and non-acupoint has some different degrees of efficacies in relieving dysmenorrhea and in upregulating spinal K-OR expression.

[Effect of electroacupuncture on the uterine microcirculation in dysmenorrhea rats].

UNLABELLED: OBJECTIVE To observe the effect of electroacupuncture (EA) on uterine microcirculation in dysmenorrhea rats so as to investigate its mechanism underlying relieving primary dysmenorrheal (PD). METHODS: Female SD rats undergoing diestrus were randomly divided into saline control, model, Sanyinjiao(SP 6), Xuanzhong(GB 39) and non-acupoint groups, with 6 rats in each. PD model was established by subcutaneous injection of estradiol benzoate for 10 days and intra-peritoneal injection of oxytocin (2 U) 1 h after the last estradiol injection. EA (2 Hz/100 Hz, 1- 1.5 mA) was applied to the above-mentioned acupoints and non-acupoint area for 20 minutes. Numbers and diameters of the uterine microvessels (11-100 microm) and capillaries (< or =10 microm), and the state of the uterine microcirculation were observed by using a Cold Light Microcirculation Detector. RESULTS: In comparison with the saline control group, the numbers and diameters of uterine microvessels and capillaries at the corresponding time-points 5 min, 10 min and 20 min were decreased significantly in the model group (P<0. 05,P<0. 01). While compared with the model group, the numbers and diameters of uterine microvessels and capillaries at 20 min after EA in the SP 6 group were increased significantly (P<0. 05). The diameter of uterus capillaries of SP 6 group was significantly bigger than that of the non-acupoint group at the time-point 20 min (P<0. 05). No significant differences were found among the SP 6, GB 39 and non-acupoint groups in the diameter of uterine microvessels (P>0. 05). CONCLUSION: EA of SP 6 can effectively increase the number of uterine microvessels and capillaries and the diameter of the uterine microvessels and capillaries in PD rats, which may contribute to its effect in relieving dysmenorrhea by improving uterine microcirculation.

[Effects of instant electroacupuncture at the different acupoints on IP3 in the uterus tissue of dysmenorrhea model rats].

OBJECTIVE: To compare the effects of instant electroacupuncture (EA) at the different acupoints on IP3 in the uterus tissue of dysmenorrhea model rats so as to investigate the specificity of acupoints. METHODS: Fifty female SD rats were randomly divided into a saline group, a model group, a Sanyinjiao (SF 6) group, a Xuehai (SP 10) group and a Hegu (LI 4) group, 10 rats in each group. The rats were given subcutaneous injection of Estradiol Beozoate injection for 10 consecutive days except those in the saline group, and intraperitoneal injection of 2U Oxytocin at 1 h after the last administration to create the dysmenorrhea rats model, and the saline group was given the same dose of saline every day. On the 10th day the rats in each EA group were given EA 20 min, and the rats in the saline group and model group were bound 20 min, and the writhing response was observed at the same time. The uterine IP3 contents were detected with enzyme-linked immunosorbent assay method. RESULTS: (1) Compared with (0.311+/- 0.253) in the saline group, the writhing scores per minute of (5.867 +/- 3.442) in the model group and (2.311 +/- 0.957) in the Xuehai (SP 10) group were both increased significantly (P < 0.01, P < 0.05), and (1.833 +/- 1.355) in the Sanyinjiao (SP 6) group and (0.743 +/- 0.306) in the Hegu (LI 4) group showed no significant differences (P > 0.05). Compared with that in the model group, the writhing scores per minute decreased significantly (all P < 0.01) in all the EA groups, with no significant differences among all the EA groups (all P > 0.05). (2) Compared with (2.698 +/- 1.491) ng/mg in the saline group, IP3 contents of the uterus of (0.813 +/- 0.899) ng/mg in the model group, (1.740 +/- 0.375) ng/mg in the Sanyinjiao (SP 6) group and (0.692 +/- 0.212) ng/mg in the Hegu (LI 4) group were all lower significantly (P < 0.05, P < 0.01), and (0.743+/- 0.306) ng/mg in the Xuehai (SP 10) group showed no significant differences (P > 0.05). Compared with that in the model group, IP3 content of the uterus in the Hegu (LI 4) group showed no significant difference (P > 0.05), and those in the Sanyinjiao (SP 6) group and in the Xuehai (SP 10) group increased significantly (both P < 0 05), which were significantly higher than that in the Hegu (II 4) group (P < 0.05, P < 0.01). CONCLUSION: There are no significant differences among the instant EA groups in improving the dysmenorrhea symptoms, but there is obvious specificity of acupoint effects in the regulation of IP3. Electroacupuncture at "Sanyinjiao (SP 6) " Xuehai (SP 10)" has more marked effect in dysmenorrhea model rats.

[Effects of electroacupuncture of "Sanyinjiao" (SP 6) on contents of MDA and beta-EP, and expression of HSP 70 in the uterus in rats with dysmenorrhea].

OBJECTIVE: To investigate the effect of electroacupuncture (EA) of "Sanyinjiao" (SP 6) on the uterus in dysmenorrhea rats so as to study its underlying analgesic mechanism. METHODS: A total of 48 SD rats during diestrus were randomized into normal saline (control) group, model group and acupuncture group according to a random number table, with 16 rats in each group. Dysmenorrhea model was established by subcutaneous injection of Estradiol benzoate (0.5 mg/d on the 1st and 10th day, and 0.2 mg/d from day 2 to day 9, once daily for 10 days) and oxytocin (2 U/rat, once on day 10). Malondialdehyde (MDA) and beta-endorphin (beta-EP) contents in the uterus were detected by radioimmunoassay, and the heat shock protein 70 (HSP 70) immunoactivity of the uterus was detected by immunohistochemistry. RESULTS: In comparison with the control group, MDA content in the uterus was increased significantly in the model group (P < 0.01), while the beta-EP level and the immunoactivity of HSP 70 immune-reaction (IR) positive products in the uterus decrease significantly (P < 0.01) and moderately, respectively in the model group. In comparison with the model group, uterine MDA content in the EA group was decreased significantly (P < 0.01), while uterine beta-EP level increased considerably (P < 0.01) and HSP 70 expression was upregulated to a certain degree. CONCLUSION: EA of "Sanyinjiao" (SP 6) can reduce MDA content and upregulate beta-EP level of the uterus in rats with dysmenorrhea, which may contribute to its analgesic effect in relieving dysmenorrhea by clearing away oxygen free radicals and raising analgesic substance in the uterus.