Effect of Modified Shaofu Zhuyutang on Neural Angiogenesis and Pelvic Pain of Endometriosis with Syndrome of Cold Congeal and Blood Stasis / 中国实验方剂学杂志

ObjectiveTo evaluate the clinical curative effect of modified Shaofu Zhuyutang on pelvic pain of endometriosis （EMT） with syndrome of cold congeal and blood stasis and the influence on neural angiogenesis. MethodA total of 110 cases were divided into a control group （54 cases） and an observation group （56 cases） by the random number table method. Patients in the control group took Aifu Nuangong pills with 6 g/time and 2 times/day. Patients in the observation group took modified Shaofu Zhuyutang with 1 dose/day. Course of treatment continued for 3 menstrual cycles. Dysmenorrhea， other symptoms and signs of pelvic pain， and the EMT health profile-5 （EHP-5） for patients with syndrome of cold congeal and blood stasis and EMT were scored before and after treatment. The levels of peripheral blood nerve growth factor （NGF）， substance P （SP）， calcitonin gene-related peptide （CGRP）， brain-derived neurotrophic factor （BDNF）， soluble Fms-like tyrosine kinase（sFlt-1）， vascular endothelial growth factor （VEGF）， matrix metalloproteinase-9 （MMP-9）， and insulin growth factor-1 （IGF-1） were detected before and after treatment. The levels of prostaglandin E2 （PGE2）， cyclooxygenase-2 （COX-2）， tumor necrosis factor-α （TNF-α）， interleukin-6 （IL-6）， and other inflammatory factors were detected before and after treatment. The pain medication usage and recurrence were recorded. ResultThe scores of dysmenorrhea， symptoms of pelvic pain symptoms （non-menstrual pelvic pain， dyspareunia， anal falling pain， and defecation pain， etc.）， and signs of pelvic pain （pelvic tenderness and sacral ligament tender nodules） in the observation group were lower than those in the control group （P<0.01）. The disappearance rates of dysmenorrhea， pelvic tenderness， and sacral ligament tender nodules were 67.35% （33/49）， 73.33% （33/45）， and 77.27% （34/44） in the observation group， which were all higher than 45.83% （22/48）， 48.84% （21/43）， and 52.27% （23/44） in the control group. The difference was statistically significant （χ2=4.571， χ2=5.565， χ2=6.026， P<0.05）. The scores of EHP-5 and syndrome of cold congeal and blood stasis in the observation group were lower than those in the control group （P<0.01）. The levels of VEGF， MMP-9， IGF-1， NGF， SP， CGRP， and BDNF in the observation group were lower than those in the control group （P<0.01）， while the level of sFlt-1 was higher than that in the control group （P<0.01）. The levels of PGE2， COX-2， TNF-α， and IL-6 in the observation group were lower than those in the control group （P<0.01）. The total effective rate of the observation group was 92.45% （49/53）， which was higher than 76.00% （38/50） of the control group （χ2=5.307， P<0.05）. After 6 months of follow-up， the recurrence rate in the observation group was 30.61% （15/49）， which was lower than 52.63% （20/38） in the control group （χ2=4.315， P<0.05）. The average of taking ibuprofen sustained-release capsules in the control group was higher than that in the observation group per menstrual period （P<0.01）. ConclusionModified Shaofu Zhuyutang treated pelvic pain of EMT with syndrome of cold congeal and blood stasis by regulating the mechanism of neural angiogenesis， reducing pain， and promoting the disappearance of related pains， thus improving the quality of life. Shaofu Zhuyutang has a better clinical effect than Aifu Nuangong pills and has a low recurrence rate.

Efficacy of Modified Huanglian Wendantang and Shaofu Zhuyutang Against Ovulation Disorder in Combined Phlegm and Stasis-Induced Polycystic Ovary Syndrome / 中国实验方剂学杂志

Objective:To observe the clinical efficacy of modified Huanglian Wendantang and Shaofu Zhuyutang in the treatment of ovulation disorder in patients with polycystic ovary syndrome （PCOS） due to the combined phlegm and stasis-induced and its influence on chronic inflammation. Method:According to the random number table， 100 patients were divided into a control group （50 cases） and an observation group （50 cases）. Apart from lifestyle intervention and oral administration of clomiphene citrate （CC） capsules to induce ovulation， patients in the control group further received Guizhi Fulingwan， 6 g/time， 2 times/day， while those in the observation group were treated with the modified Huanglian Wendantang and Shaofu Zhuyutang， 1 dose/day， for six menstrual cycles. The ovulation， endometrial thickness， proportion of type A endometrium， as well as the pulsatility index （PI） and resistance index （RI） of uterine artery were monitored before and after treatment. The fasting blood glucose （FBG）， fasting insulin （FINS）， luteinizing hormone （LH）， follicle stimulating hormone （FSH）， serum testosterone （T）， estradiol （E₂）， dehydroepiandrosterone sulfate （DHEAS）， tumor necrosis factor-<italic>α</italic> （TNF-<italic>α</italic>）， high-sensitivity C-reactive protein （hs-CRP）， adiponectin （APN）， and interleukin-6 （IL-6） levels before and after treatment were detected， followed by the calculation of homeostasis model assessment-insulin resistance （HOMA-IR） value and the evaluation of ovarian volume and combined phlegm and stasis-induced syndrome score. Result:The overall response rate of the observation group was （44/47） 93.62%， which was higher than （36/46） 78.26% of the control group （<italic>χ</italic>²=4.802， <italic>P</italic><0.05）. The ovulation rate in the observation group was （199/264） 75.38%， higher than （173/272） 63.60% in the control group （<italic>χ</italic>²=8.714， <italic>P</italic><0.01）. The clinical pregnancy rate of the observation group was （11/47） 23.40%， higher than （5/46） 10.87% of the control group， but the difference was not statistically significant （<italic>χ</italic>²=2.564， <italic>P</italic>>0.05）. Compared with the control group， the observation group exhibited reduced PI， RI， LH， T， DHEAS， FINS， FPG， HOMA-IR， TNF-<italic>α</italic>， hs-CRP， and IL-6 （<italic>P</italic><0.01）， but elevated E₂， FSH， and APN （<italic>P</italic><0.01）. Besides， the bilateral ovarian volume and combined phlegm and stasis-induced syndrome score of the observation group were smaller than those of the control group （<italic>P</italic><0.01）， while the endometrial thickness and proportion of type A endometrium were higher （<italic>P</italic><0.01）. Conclusion:On the basis of CC treatment， the modified Huanglian Wendantang and Shaofu Zhuyutang alleviates the ovulation disorder in PCOS patients of combined phlegm and stasis-induced syndrome and regulates IR and chronic inflammation， thus creating a favorable condition for clinical pregnancy， which is worthy of further research.

Effect and Mechanism of Shaofu Zhuyutang on Endothelial Injury of Rats with Cold Coagulation and Blood Stasis Syndrome Based on Nrf2/ARE Signaling Pathway / 中国实验方剂学杂志

Objective:To observe the effect of Shaofu Zhuyutang on nuclear factor erythroid-2-related factor 2 （Nrf2） /antioxidant response element （ARE） signaling pathway in blood vessels by establishing the model of rats with cold coagulation and blood stasis syndrome， and to explore the protective effect and mechanism of Shaofu Zhuyutang on vascular endothelial injury. Method:The 50 SPF rats were randomly divided into high dose group （4.8 g·kg-1）， middle dose group （2.4 g·kg-1）， low dose group （1.2 g·kg-1）， model group and normal group （ten of each group）. The rat model of cold coagulation and blood stasis syndrome was established by subcutaneous injection of epinephrine hydrochloride combined with ice bath. At the same time of modeling， the drug was administered by gavage. After 28 days of continuous administration， the hemorheology indexes were detected by automatic hemorheology instrument. Levels of nitric oxide （NO）， endothelin （ET）-1， superoxide dismutase （SOD）， glutathione （GSH-Px）， intercellular adhesion molecule-1（ICAM-1）， vascular cell adhesion molecule-1（VCAM-1）， von Willebrand factor （vWF） in serum were determined by ELISA. Hematoxylin and eosin （HE） staining was used to observe the endothelial injury of vascular tissue of thoracic aorta. The protein expression of Nrf2 and HO-1 in vascular tissue of thoracic aorta was detected by Western blot. Real-time fluorescence quantitative polymerase chain reaction（Real-time PCR）was used to observe the expression of Nrf2 and heme oxygenase-1 （HO-1） mRNA in vascular tissue of thoracic aorta. Result:Compared with the blank group， model group rats whole blood viscosity and plasma viscosity were significantly increased （P<0.05，P<0.01）， vWF， ICAM 1， VCAM 1 content increased significantly （P<0.01）， NO， SOD， gsh-px levels decreased significantly （P<0.01）， significantly increased the content of ET-1（P<0.01）， thoracic aorta vascular tissue Nrf2， HO-1 mRNA expression was significantly increased （P<0.01）， Nrf2 protein expression in the cell nucleus increased significantly （P<0.05）， The protein expression level of Nrf2 in cytoplasm was significantly decreased （P<0.05）， while the protein expression level of HO-1 was significantly increased （P<0.01）. Compared with model group， the whole blood viscosity （high and middle cut）， plasma viscosity， were significantly reduced in high and meduim-dose Shaofu Zhuyutang groups（P<0.05，P<0.01）. The levels of vWF， ICAM-1， VCAM-1 and ET-1 in serum were significantly reduced （P<0.05，P<0.01）， NO， SOD and GSH-Px increased significantly （P<0.05，P<0.01）. The pathological changes such as hyperplasia， swelling and shedding of endothelial cells of thoracic aorta， rupture of internal elastic membrane and disorder of smooth muscle arrangement were improved. The expression levels of Nrf2， HO-1 protein and gene were significantly increased in vascular tissue of thoracic aorta （P<0.01）. Conclusion:Shaofu Zhuyutang has a protective effect on vascular endothelial injury in rats with cold coagulation and blood stasis syndrome. The mechanism of action is related to the activation of Nrf2/ARE signaling pathway， which leading to the increased expression of antioxidant enzymes and decreased the expression of adhesion factors.

Clinical Efficacy of Modified Shaofu Zhuyutang Combined with Zhengs Rebuzhen Therapy on Pain Caused by Adenomyosis / 中国实验方剂学杂志

Objective:To observe the effect of modified Shaofu Zhuyutang combined with Zheng's Rebuzhen therapy on pain caused by adenomyosis (cold coagulation and blood stasis syndrome) and levels of vascular endothelial growth factor (VEGF), matrix metalloproteinases-9 (MMP-9), tissue inhibitors of matrix metalloproteinases (TIMP-1) and prostaglandin. Method:One hundred and twenty-eight patients were randomly divided into control group (63 cases) and observation group (65 cases) by random number table. Patients in control group got desogestrel and ethinylestradiol tablets since the first day of menstruation for consecutive 21 days, 1 granule/time, 1 time/day, and Zheng's Rebuzhen therapy for consecutive 10 days during the menstruation, 1 time/day. In addition to the therapy of control group, patients in observation group were also given Shaofu Zhuyutang, 1 dose/day. A course of treatment was 3 menstrual cycles. Before and after treatment, cox menstrual symptom scale (CMSS) of COX, visual simulation of dysmenorrheal (VAS), chronic pelvic pain, pictorial blood loss assessment chart (PBAC) and cold coagulation and blood stasis syndrome were scored. And uterine volume and endometrial thickness were evaluated by B-mode ultrasonography. And levels of VEGF, MMP-9, TIMP-1, prostaglandin E2 (PGE2), prostaglandin F2α(PGF2α) and cancer antigen were detected. Result:After treatment, the clinical efficacy in observation group was superior to that in control group (Z=2.445, PPP2α and CA125 were lower than those in control group (P2 were higher than those in control group (PConclusion:In addition to desogestrel and ethinylestradiol tablets combined with Zheng's Rebuzhen therapy, modified Shaofu Zhuyutang can relieve pain and hemorrhage and improve the clinical efficacy. The mechanism may be correlated with the regulation of expression levels of VEGF, MMP-9, TIMP-1 and prostaglandin.