[Effect of Liangfang Wenjing Decoction on expression of key glycolytic enzymes in uterus and ovaries of rats with coagulating cold and blood stasis syndrome].

This study aimed to investigate the relationship between coagulating cold and blood stasis syndrome and glycolysis, and observe the intervention effect of Liangfang Wenjing Decoction(LFWJD) on the expression of key glycolytic enzymes in the uterus and ovaries of rats with coagulating cold and blood stasis. The rat model of coagulating cold and blood stasis syndrome was established by ice-water bath. After modeling, the quantitative scoring of symptoms were performed, and according to the scoring results, the rats were randomly divided into a model group and LFWJD low-, medium-and high-dose groups(4.7, 9.4, 18.8 g·kg~(-1)·d~(-1)), with 10 in each group. Another 10 rats were selected as the blank group. After 4 weeks of continuous administration by gavage, the quantitative scoring of symptoms was repeated. Laser speckle flowgraphy was used to detect the changes of microcirculation in the ears and uterus of rats in each group. Hematoxylin-eosin(HE) staining was used to observe the pathological morphology of uterus and ovaries of rats in each group. The mRNA and protein expressions of pyruvate dehydrogenase kinase 1(PDK1), hexokinase 2(HK2) and lactate dehydrogenase A(LDHA) in the uterus and ovaries of rats were examined by real-time quantitative polymerase chain reaction(RT-qPCR) and Western blot, respectively. The rats in the model group showed signs of coagulating cold and blood stasis syndrome, such as curl-up, less movement, thickened veins under the tongue, and reduced blood perfusion in the microcirculation of the ears and uterus, and HE staining revealed a thinning of the endometrium with disorganized arrangement of epithelial cells and a decrease in the number of ovarian follicles. Compared with the model group, the treatment groups had alleviated coagulating cold and blood stasis, which was manifested as red tongue, reduced nail swelling, no blood stasis at the tail end as well as increased blood perfusion of the microcirculation in the ears and uterus(P<0.05 or P<0.01). Among the groups, the LFWJD medium-and high-dose groups had the most significant improvement in coagulating cold and blood stasis, with neatly arranged columnar epithelial cells in uterus, and the number of ovarian follicles was higher than that in the model group, especially mature follicles. The mRNA and protein expressions of PDK1, HK2, LDHA in uterus and ovaries were up-regulated in the model group(P<0.05 or P<0.01), while down-regulated in LFWJD medium-and high-dose groups(P<0.05 or P<0.01). The LFWJD low-dose group presented a decrease in the mRNA expressions of PDK1, HK2 and LDHA in uterus and ovaries as well as in the protein expressions of HK2 and LDHA in uterus and HK2 and PDK1 in ovaries(P<0.05 or P<0.01). The therapeutic mechanism of LFWJD against coagulating cold and blood stasis syndrome is related to the down-regulation of key glycolytic enzymes PDK1, HK2 and LDHA, and the inhibition of glycolytic activities in uterus and ovaries.

Screening and Identification of the Main Metabolites of Schisantherin a In Vivo and In Vitro by Using UHPLC-Q-TOF-MS/MS.

Schisantherin A is an active ingredient originating from Schisandra chinensis (Turcz.) which has hepatoprotective and anti-oxidation activities. In this study, in vitro metabolisms investigated on rat liver microsomes (RLMs) and in vivo metabolisms explored on male Sprague Dawley rats of Schisantherin A were tested, respectively. The metabolites of Schisantherin A were identified using ultra-high-performance liquid chromatography coupled with hybrid triple quadrupole time-of-flight mass spectrometry (UHPLC-Q-TOF-MS/MS). Based on the method, 60 metabolites were successfully identified and structurally characterized including 48 phase-I and 12 phase-II metabolites. Among the metabolites, 45 metabolites were reported for the first time. Moreover, 56 and eight metabolites were detected in urine and bile and 19 metabolites were identified in rats' plasma. It demonstrated that hepatic and extra-hepatic metabolic pathways were both involved in Schisantherin A biotransformation in rats. Five in vitro metabolites were structurally characterized for the first time. The results indicated that the metabolic pathways mainly include oxidation, reduction, methylation, and conjugation with glucuronide, taurine, glucose, and glutathione groups. This study provides a practical strategy for rapidly screening and identifying metabolites, and the results provide basic data for future pharmacological and toxicology studies of Schisantherin A and other lignin ingredients.

[Effect of Acupoint Application Therapy of Summer Treatment for Winter Diseases on Nerve-Endocrine-Immune Network System in Patient with Non-Acute Attack Asthma].

Objective To observe the effect of acupoint application therapy in summer to treat winter diseases (abbreviated as AATSTWD) on nerve-endocrine-immune network system in patients with non-acute attack asthma, and to study possible mechanisms. Methods Fifty healthy volunteers were re- cruited as the normal control group and 50 patients with non-acute attack asthma were recruited as the asthma group. Patients in the normal control group received no intervention. Those in the control group were treated with acupoint application therapy on hot days (applied on the first day of three dog days, 3 times in total). Blood samples were tested before treatment and after 3 times of application. The contents of serum immunoglobulin E (IgE) , interleukin-4 (IL-4) , plasma substance P (SP) , and vasoactive intes- tinal peptide (VIP) were detected using radioimmunoassay. Contents of immunoglobulin A and G (IgA, IgG) were tested by immunoturbidimetry. Content of interferon-y (INF-y) were tested by enzyme linked immunosorbent assay. Results Compared with the normal control group, serum contents of IgA, igG, IFN-y, and plasma VIP decreased, contents of IgE, IL-4, and SP significantly increased in the asthma group before treatment (P <0. 05). Compared with before treatment in the same group, serum contents of IgA, IgG, VIP, and IFN-y increased, and contents of IgE, IL-4, and SP decreased in the asthma group after treatment (P <0. 05). Conclusion Acupoint application therapy in summer to treat winter diseases method could prevent and treat bronchial asthma possibly through improving immune function, control- ling the release of cytokines , and regulating neurotransmitters.

Clerodens E-J, antibacterial caffeic acid derivatives from the aerial part of Clerodendranthus spicatus.

Six new caffeic acid derivatives, Clerodens E-J (1-6) were isolated from the aerial part of Clerodendranthus spicatus. Their structures were elucidated by extensive spectroscopic analysis, including NMR, MS, and ECD data. Compound 1 showed moderate antibacterial activities against drug-resistant strains of bacteria in vitro.

[Effect of electroacupuncture intervention at different time-points of post-modeling on hippocampal monoamine neurotransmitter levels in mice with vascular dementia].

OBJECTIVE: To observe the effect of electrocupuncture (EA) intervention at different time-points of post-modeling on behavior and hippocampal monoamine neurotransmitter noradrenalin (NE), dopamine (DA) and 5-hydroxytrypamine (5-HT) contents in vascular dementia (VD) mice, so as to study its mechanism underlying improvement of VD. METHODS: A total of 60 Kunming mice were randomized into sham-operation control (n = 20), VD model (n = 20), EA-day (D)-1 (EA treatment was given from the 1st day on after modeling, n = 10), EA-D-3 (EA was given from the 3rd day on after modeling, n = 10) groups. VD model was established by occlusion of the bilateral cervical common arteries and reperfusion. EA (2 Hz/80 Hz) was applied to "Baihui" (GV 20), "Dazhui" (GV 14),"Zusanli" (ST 36) and "Geshu"(BL 17) for 10 min, once daily for 15 days. Hippocampal NE, DA and 5-HT contents were assayed by fluorospectrophotometry. The mouse's learning-memory ability was assessed by step-down tests. RESULTS: In comparison with the sham-operation control group, the learning-memory ability (marked increase of reaction time and error times, decrease of step-down latency) was apparently lowered in the model group (P < 0.01). The hippocampal NE, DA and 5-HT contents were significantly lower in the model group than in the sham-operation group (P < 0.01). Compared with the model group, the mice's learning-memory ability (marked decrease of reaction time and error times, increase of step-down latency) was significantly increased in EA intervention groups (P < 0.01), and hippocampal NE, DA and 5-HT levels were significantly increased (P < 0.01), and the effect of EA-D-3 group was obviously better than that of the EA-D-1 group (P < 0.05, P < 0.01). CONCLUSION: EA can improve the VD mice's learning-memory ability, which is closely related to its effects in up-regulating hippocampal NE, DA and 5-HT contents, and the effect of later EA intervention after modeling is better.

[Effect of gufuhuo decoction on local apoptosis of steroid-induced necrosis of femoral head].

OBJECTIVE: To observe the bone specimen of SANFH (steroid-induced avascular necrosis of femoral head) and the effect of Gufuhuo decoction on local apoptosis of SANFH. METHOD: Model rabbits of necrosis of femoral head, made by intragluteal injection with prednisone acetate, were killed after 6 weeks to study the apoptosis by means of TUNEL. RESULT: A great number of faded osteocytes and osteoblasts existed in specimen of SANFH. There was significant difference in apoptosis indexes between model and control group, Gufuhuo decoction and Xianlinggubao group (P < 0.05). CONCLUSION: In the course of SANFH, the necrosis results from the interaction of apoptosis and necrosis of steroid-induced osteocytes and osteoblasts. Gufuhuo decoction has some effects on improving apoptosis and necrosis of SANFH.

[Effect of gufuhuotang on hemorheology and lipid metabolism of hormonal necrosis of femoral head].

OBJECTIVE: To probe the of Gufuhuotang effect on hemorheology and lipid metabolism of hormonal necrosis of femoral head. METHOD: 32 male rabbits of New Zealand were divided at random into pathological pattern group, normal control group, low-dosage of Gufuhuotang group and high-dosage group; models of osteocyte necrosis of head of femur were made with intragluteal injection of hydroprednisone acetate; the rabbits were observed weekly and killed 6 weeks later and HE pathological section was made to observe hemorheology index and blood-liquid content, and then analysis and comparison were made. RESULT: Gufuhuotang could obviously decrease total blood low-shear viscosity, plasma viscosity, hemacocrit, serum cholesterol and triglyceride contents; under light microscope empty bone lacunas of femoral head were less, fat cells of medullary cavity were less and relatively small, and bone trabecula was sparse with no break. CONCLUSION: Gufuhuotang has a distinct preventive and curative effect on rabbits' hormonal necrosis of femoral head.