Effect and Underlying Mechanism of Bu-Shen-An-Tai Recipe on Ovarian Apoptosis in Mice with Controlled Ovarian Hyperstimulation Implantation Dysfunction

The effect and underlying mechanism of Bu-Shen-An-Tai recipe on ovarian apoptosis in mice with controlled ovarian hyperstimulation(COH) implantation dysfunction were studied. The COH implantation dysfunction model in mice was established by intraperitoneal injection of 7.5 IU pregnant mare’s serum gonadotrophin(PMSG), followed by 7.5 IU human chorionic gonadotrophin(HCG) 48 h later. Then the female mice were mated with male at a ratio of 2:1 in the same cage at 6:00 p.m. The female mice from normal group were injected intraperitoneally with normal saline and mated at the corresponding time. Day 1 of pregnancy was recorded by examining its vaginal smears at 8:00 a.m. of the next day. Fifty successfully pregnant mice were equally randomly divided into 5 groups: normal control pregnant group(NC), COH implantation dysfunction model group(COH), low dosage of Bu-Shen-An-Tai recipe group(LOW), middle dosage of Bu-Shen-An-Tai recipe group(MID) and high dosage of Bu-Shen-An-Tai recipe group(HIGH). Then from day 1, the mice in different groups were respectively intragastrically given corresponding treatments at 9:00 a.m. for 5 consecutive days. The concentrations of 17β-estradiol(E) and progesterone(P) were determined by radioimmunoassay(RIA). The ultrastructural changes of ovarian tissues were observed by transmission electron microscope(TEM). The histopathological changes of ovarian tissues were observed by HE staining. The number of atretic follicles and pregnant corpus luteum were also recorded. TUNEL was applied to measure apoptotic cells of ovarian tissues. Western blotting was used to detect the protein expression of apoptosis-related factors like Bax, Bcl-2 and cleaved-caspase-3 in ovarian tissue of mice. The results showed that ovarian weight, the concentrations of Eand P, the number of atretic follicles and pregnant corpus luteum, as well as the apoptosis of granulosa cells were significantly increased in the COH group. The ultrastructures of ovarian tissues in the COH group showed that chromatin in granulosa cells was increased, agglutinated, aggregated or crescent-shaped. The focal cavitation and the typical apoptotic bodies could be seen in granulosa cells in the late stage of apoptosis. After the treatment with different doses of Bu-Shen-An-Tai recipe, the ultrastructural changes of ovarian granulosa cells apoptosis were dramatically improved and even disappeared under TEM. Visible mitochondria and mitochondrial cristae were increased and vacuoles were significantly reduced. The lipid dropltes were shown in a circluar or oval shape. The protein expression levels of Bax and cleaved-caspase-3 were decreased, and the expression of Bcl-2 protein was increased after treatment. It was concluded that Bu-Shen-An-Tai recipe can inhibit the apoptosis of ovarian granulosa cells, probably by up-regulating the protein expression of Bcl-2 and down-regulating Bax and cleaved-caspase-3, which contributes to the formation and maintenance of ovarian corpus luteum. It’s helpful to promote the embryonic implantation, to reduce embryo loss and ultimately to improve the success rate of pregnancy.

Evaluation of Biochemical and Molecular Parameters of Patients Suffering from Chronic Viral Hepatitis B Treated by a Medicinal Plant Recipe of a Health Care Practitioner in Burkina Faso

Chronic viral hepatitis B (HBV) remains a major public health problem in Burkina Faso. Since access to diagnostic tests and treatments is limited because of their high cost, the majority of the population turn to traditional herbal treatments. This study aimed to evaluate the effectiveness of a plant recipe called Hepatib tiben. It consisted of comparing certain biochemical and molecular parameters of patients infected with HBV that were supported by the recipe. The patients were recruited in Ouagadougou by the traditional health practitioner according to the requirements of the study. Thus 44 patients aged 20 to 61 years and carrier of HBsAg for at least 06 months were treated with Hepatib tiben. The tests were performed in the laboratory before and three months after the treatment. ELISA tests were used to confirm the presence of HBsAg and search for anti-HCV antibodies;transaminases, creatinine were quantified by the “Chem 400” automaton and the viral load of HBV by Real-time PCR. The analysis of the results reveals an improvement of the biochemical and molecular parameters of the patients with the following means (ASAT: 21.02 ± 9.97;ALAT: 21.11 ± 13.27;DNA: 1571.82 ± 3990.97 with p = 0.01 for each). As for HBsAg, its disappearance was observed in 4.55% of patients after treatment. The evaluation of the creatinine parameter explained that the recipe of plants has a tolerated effect on the kidneys of treated patients. These results, while encouraging, need to be complemented by further research for the development of effective phytomedicine to treat and eliminate this viral hepatitis B virus.

Exploring the effect of Bushen Bitong recipe-containing serum on IL-1β-induced chondrocyte apoptosis based on SOX9/NF-κB/MMP-13 signaling pathway

Objective:To observe the effect and possible mechanism of action of Bushen Bitong recipe(BSBT)containing serum on IL-1β-induced chondrocyte apoptosis.Methods:Generation 3 rat chondrocytes were randomized into Control,IL-1β,IL-1β+BSBT(L),IL-1β+BSBT(M),and IL-1β+BSBT(H)groups(5%,10%and 15%BSBT-containing serum),and then 24h after intervention respectively,the cell proliferation and Apoptosis rate;Western blot detected the expression levels of Bcl-2,BAX,Caspase-3,SOX9,NF-κB p65,MMP-13 proteins in chondrocytes.ELISA detected the levels of TNF-α,IL-6,and bFGF in the supernatants of chondrocyte culture.Results:Compared with Control group,cell proliferation activity decreased,apoptosis rate increased,NF-κB p65,MMP-13 protein level and TNF-α,IL-6 level increased,and SOX9 protein level and bFGF level decreased in IL-1βgroup;compared with IL-1βgroup,different concentrations of BSBT-containing serum group,cell proliferation activity increased,and apoptosis rate decreased.NF-κB p65,MMP-13 protein level and TNF-α,IL-6 level decreased,SOX9 protein level and bFGF level increased;compared with IL-1β+BSBT(L)group,cell proliferation activity increased,apoptosis rate decreased in IL-1β+BSBT(M)and IL-1β+BSBT(H)groups,and NF-κB p65,MMP-13 protein level and TNF-αlevel decreased.13 protein levels and TNF-αand IL-6 levels decreased,and SOX9 protein levels and bFGF levels increased.Conclusion:BSBT-containing serum may promote IL-1β-induced proliferation of chondrocytes,reduce apoptosis,improve the microenvironment of chondrocytes,and promote cartilage repair through the SOX9/NF-κB/MMP-13 signaling pathway.

Exploring the therapeutic potential of Qi Teng Mai Ning recipe in ischemic stroke and vascular cognitive impairment

Background:This study aims to explore the therapeutic effects of the Qi Teng Mai Ning recipe on ischemic stroke and vascular cognitive impairment through its potential to modulate cellular autophagy,with a focus on identifying its active ingredients and their target proteins.Methods:The study began with the identification of active ingredients in the Qi Teng Mai Ning recipe.It proceeded to screen the gene expression omnibus database for ischemic stroke and vascular cognitive impairment-associated differentially expressed mRNAs and to identify cellular autophagy-related proteins via the Human Autophagy Database.These proteins were annotated with Gene Ontology and Kyoto Encyclopedia of Genes and Genomes functions and subjected to molecular docking with the recipe’s core active ingredients.In vitro cell experiments were conducted on hypoxic HT22 cells,involving CCK8 assay,lentiviral transfection to silence autophagy related 9B(ATG9B),immunofluorescence staining,and qPCR validation to investigate the effects of the recipe on autophagy.Results:The analysis identified 104 active ingredients targeting 408 proteins and forming a complex ingredient-target network.Intersecting 55 ischemic stroke-related and 909 vascular cognitive impairment-related differentially expressed mRNAs revealed 14 co-expressed mRNAs.Molecular docking showed quercetin,kaempferol,myrcene,and conferone as key ingredients targeting autophagy-related proteins.Cellular experiments indicated that the recipe significantly enhanced cell viability under hypoxic conditions,reduced apoptosis,and modulated the expression of autophagy-related factors,thereby decreasing apoptosis rates in HT22 cells.Conclusion:The Qi Teng Mai Ning recipe offers protective effects against ischemic stroke and vascular cognitive impairment by modulating autophagy-related proteins.Its efficacy highlights the potential of traditional Chinese medicine in treating these conditions,though further research is needed to fully understand its mechanisms and clinical applications.

Therapeutic Effect of Intragastric Administration of Jiangtang Shuxin Recipe Suspension on Diabetic Heart Failure in Rats and Its Mechanism

[Objectives]To observe the therapeutic effect of intragastric administration of Jiangtang Shuxin recipe on diabetic heart failure(DHF)in rats and to explore its mechanism.[Methods]Fifty SD rats were randomly divided into five groups,with 10 rats in each group.DHF models were prepared in the low-dose group,high-dose group,Western medicine group,and model group except the control group.Rats in the low-dose and high-dose groups were given 1.0 and 1.5 g/(kg·d)Jiangtang Shuxin recipe suspension by gavage,respectively.Rats in the Western medicine group were given gliquidone and benazepril by gavage for 2 months,and were fed with high-fat diet.Rats in the control group were fed with ordinary diet.Fasting blood glucose(FBG),serum triglyceride(TG),low-density lipoprotein cholesterol(LDL-C),high-density lipoprotein cholesterol(HDL-C),C-reactive protein(CRP),interleukin-6(IL-6),tumor necrosis factor-α(TNF-α),creatine kinase isoenzyme(CK-MB),lactate dehydrogenase(LDH),pathological morphology of myocardial tissue,NF-κB p65 protein and IκBαprotein were compared among groups.[Results]Compared with the control group,the levels of FBG,serum TG,LDL-C,CRP,IL-6,TNF-α,CK-MB and LDH increased,while the level of serum HDL-C decreased.The myocardial tissue was seriously damaged,and the expression of NF-κB p65 protein increased,while the expression of IκBαprotein decreased in the other four gruops(all P<0.05).Compared with the model group,the levels of FBG,serum TG,LDL-C,CRP,IL-6,TNF-α,CK-MB and LDH decreased,while the serum HDL-C level increased.The myocardial tissue damage was alleviated,and the expression of NF-κB p65 protein decreased,while the expression of IκBαprotein increased in the low-dose group,high-dose group and Western medicine group(all P<0.05).Compared with the Western medicine group,the levels of FBG,serum TG,LDL-C,CRP,IL-6,TNF-α,CK-MB and LDH decreased,and the level of serum HDL-C increased in the high-dose group(all P<0.05).[Conclusions]Jiangtang Shuxin recipe has a therapeutic effect on DHF in rats,with the best effect in the high-dose group.It can not only alleviate high glucose and high fat state,but also reduce myocardial injury and inflammation,and improve the pathological morphology of myocardial cells.The mechanism may be related to its inhibition of NF-кB signaling pathway.

Network pharmacology-based exploration of molecular mechanisms underlying therapeutic effects of Jianpi Huatan Quyu recipe on chronic heart failure with spleen Qi deficiency syndrome

BACKGROUND Chronic heart failure is a complex clinical syndrome.The Chinese herbal compound preparation Jianpi Huatan Quyu recipe has been used to treat chronic heart failure;however,the underlying molecular mechanism is still not clear.AIM To identify the effective active ingredients of Jianpi Huatan Quyu recipe and explore its molecular mechanism in the treatment of chronic heart failure.METHODS The effective active ingredients of eight herbs composing Jianpi Huatan Quyu recipe were identified using the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform.The target genes of chronic heart failure were searched in the Genecards database.The target proteins of active ingredients were mapped to chronic heart failure target genes to obtain the common drugdisease targets,which were then used to construct a key chemical componenttarget network using Cytoscape 3.7.2 software.The protein-protein interaction network was constructed using the String database.Gene Ontology and Kyoto Encyclopedia of Genes and Genomes enrichment analyses were performed through the Metascape database.Finally,our previously published relevant articles were searched to verify the results obtained via network pharmacology.RESULTS A total of 227 effective active ingredients for Jianpi Huatan Quyu recipe were identified,of which quercetin,kaempferol,7-methoxy-2-methyl isoflavone,formononetin,and isorhamnetin may be key active ingredients and involved in the therapeutic effects of TCM by acting on STAT3,MAPK3,AKT1,JUN,MAPK1,TP53,TNF,HSP90AA1,p65,MAPK8,MAPK14,IL6,EGFR,EDN1,FOS,and other proteins.The pathways identified by KEGG enrichment analysis include pathways in cancer,IL-17 signaling pathway,PI3K-Akt signaling pathway,HIF-1 signaling pathway,calcium signaling pathway,cAMP signaling pathway,NF-kappaB signaling pathway,AMPK signaling pathway,etc.Previous studies on Jianpi Huatan Quyu recipe suggested that this Chinese compound preparation can regulate the TNF-α,IL-6,MAPK,cAMP,and AMPK pathways to affect the mitochondrial structure of myocardial cells,oxidative stress,and energy metabolism,thus achieving the therapeutic effects on chronic heart failure.CONCLUSION The Chinese medicine compound preparation Jianpi Huatan Quyu recipe exerts therapeutic effects on chronic heart failure possibly by influencing the mitochondrial structure of cardiomyocytes,oxidative stress,energy metabolism,and other processes.Future studies are warranted to investigate the role of the IL-17 signaling pathway,PI3K-Akt signaling pathway,HIF-1 signaling pathway,and other pathways in mediating the therapeutic effects of Jianpi Huatan Quyu recipe on chronic heart failure.

Pharmacokinetics of traditional Chinese syndrome and recipe:a hypothesis and its verification(Ⅰ)

AIM To propose a hypothesis defining theabsorption,distribution,metabolism andelimination of traditional Chinese recipe（TCR）-component in blood of healthy subjects andpatients,and estimate its correctness.METHODS The pharmacokinetics（PK）of samedose of drug was studied in the animal model oftraditional Chinese syndrome（S）and healthyanimals.The classification,terminology,concept and significance of the hypothesis wereset forth with evidence provided in the presentstudy.The hypotheses consisted of traditionalChinese syndrome PK（S-PK）and traditionalChinese recipe PK（R-PK）.Firstly,the observedtetramethylpyrazine（TMP）PK in healthy,chronically reserpinized rats（rat model ofspleen deficiency syndrome,RMSDS）andRMSDS treated with Sijunzi decoction（SJZD）forconfirmation were used to verify S-PK; secondly,the ferulic acid（FA）PK in healthy andhigh molecular weight dextran（HMWD）-inducedrabbit model with blood stasis syndrome（RDBSS）was also used to verify S-PK;andlastly,TMP PK parameters in serum of healthyrats after orally taken Ligusticum wallichii（LW）,LW and Salvia miltiorrhiza（LW&SM）decoctions were compared to verify R-PK.RESULTS The apparent first-order absorption[Ka,（13.61±2.56）h-1,area under the blooddrug concentration-time curve[AUC,（24.88±9.76）μg.h-1mL-1],maximum drug concentration[Cmax,（4.82±1.23）μg·mL-1]of serum TMP inRMSDS were increased markedly（P【0.05）compared with those[Ka=（5.41±1.91）h-1,AUC=（5.20±2.57）μg·h-1·mL-1,Cmax=（2.33±1.77）μg·mL-1]of healthy rats（HR）.Theapparent first-order rate constant for α and βdistribution phase[α=（0.38±0.09）h-1,β=（0.06±0.03）h-1,the apparent first-orderintercompartmental transfer rate constants[K10=（0.24±0.07）h-1,K12=（0.11±0.02）h-1,K21=（0.11±0.02）h-1]of serum TMP in RMSDS weredecreased significantly（P【0.01）comparedwith those[K10=（0.88±0.20）h-1,K12=（1.45±0.47）h-1,K21=（0.72±0.22）h-1]of HR.However,no apparent differences occurredbetween HR and RMSDS treated with SJZD.Theserum FA concentration and its AUC[（5.6690±2.3541）μg·h-1·mL-1] in RMBSS were also higherthan those[AUC=（2.7566±0.8232）μg·h-1·mL-1]of healthy rabbits（P【0.05）.The Ka（11.51±2.82）h-1,AUC（0.84±0.17）μg·h-1·mL-1of LW & SM-derived TMP in serum weremuch lower（P【0.05）than those[Ka=（19.58±4.14）h-1,AUC=（1.27±0.26）μg·h-1·mL-1]ofLW-derived TMP in serum after oral decoctions.CONCLUSION The SDS and blood stasissyndrome state could affect significantly thepharmacokinetic parameters of drugs and theabnormal SDS pharmacokinetic parameters couldbe normalized by SJZD.The combination ofChinese medicine in TCR could reciprocallyaffect the pharmacokinetic parameters of othercomponents absorbed into the systemiccirculation.These results support the S-and R-PK hypothesis.

Effect of Liangxuehuayu Recipe on hemorheology in rats with blood stasis syndrome

Objective:To investigate the effects of Uangxuehuayu Recipe on hemorheology in rats with blood stasis syndrome induced by mutifactor stimuli.Methods:SD rats were divided into control, model,Uangxuehuayu Recipe(high,middle and low dose,18,9,4.S g/kg accordingly).Except the control group,blood stasis model was established in the rest groups.The hemorheological parameters were measured and compared.Results:Blood viscosity at high,moderate and low level in rats with blood stasis significantly increased(P【0.05),but blood viscosity at high level and plasma viscosity was significantly decreased in rats induced by some stimuli after Uangxuehuayu Recipe were intra-gastrically administered for I weeks(P【0.01,P【0.05). Conclusions:Uangxuehuayu Recipe is effective in improving hemorheology,and has important application value in the prevention of occurrence and development of ischemic stroke.

Two-year Observation of the Clinical Efficacy in Treating Chronic Hepatitis B Patients with Ganxian Recipe (肝纤方) and Lamivudine

Objective: To evaluate the clinical efficacy of Ganxian recipe (肝纤方, GXR) and lamivudine (LVD) in a two-year treatment of chronic hepatitis B (CHB). Methods: One hundred and twenty patients with CHB were randomly divided into the combinedly treated group (combined group) of 40 CHB patients who were treated with GXR combined with LVD. Another 40 CHB patients were treated with LVD alone (WM group), and still another 40 CHB patients were treated with GXR alone (TCM group). All these cases were randomly controlled and observed for two years. Results: Comprehensive efficacy: Total effective rate of the combined group (complete response and partial response) was 92.5%, while that of the WM group was 67.5% and TCM group 57.5%, respectively, with the difference between them was significant ( P <0.01); after treatment, the hepatic functions (AST, ALT, SB) of the three groups were all reduced, and the reduction in the combined group was particularly significant in comparison with the WM group or TCM group, P <0.05 or P < 0.01 respectively, suggesting that the effect in the combined group was better than that in the other two groups; the rate of tyrosine-methionine-aspartate-aspartate (YMDD) virus mutation: it was 7.5% in the combined group, 40.0% in the WM group, and 5.0% in the TCM group; liver fibrosis improvement parameter: after treatment, the results in the combined group got better than those in the other two groups. Conclusion: GXR could inhibit the appearance of YMDD after long-term application of LVD, and combined use has marked synergism.

Effect of Gengnianchun Recipe (更年春方) on Bone Mineral Density, Bone Biomechanical Parameters and Serum Lipid Level in Ovariectomized Rats

Objective： To observe the effect of Gengnianchun Recipe （更年春方, GNC） on bone mineral density （BMD）, bone biomechanical parameters and serum lipid level in the bilaterally ovariectomized （OVX） rats and to explore the prophylactic and therapeutic action of GNC on ovariectomy induced osteoporosis and hyperlipidemia. Methods： OVX SD rats, 10- 12 months old, were divided into different groups and fed with GNC 2 g/d, GNC 1 g/d and Nilestriol 0. 125 mg/week, respectively for 4 months to observe the change of BMD and bone biomechanical parameters of the lumbar vertebrae, and the serum levels of total cholesterol （TO）, triglyceride（TG）, high-density lipoprotein cholesterol （HDL-C） and low-density lipoprotein cholesterol （LDL-C）, and to compare the effect of the two drugs on the morphology of the uterus. Results： There was marked reduction in BMD and biomechanical parameters in lumbar vertebrae （ P〈0. 01 ） and increase of serum TO and LDL-C levels （ P〈0. 01 ） in rats after OVX. GNC or Nilestriol significantly improved the decreased BMD and biomechanical parameters of the lumbar vertebrae （ P〈0.05 or P〈0. 01 ）, and reduced the serum TO and LDL-C levels （ P〈0. 01 ）. In the Nilestriol group, the wet weight of uterus got increased obviously （ P〈0.01 ）, the number of uterine glands increased, uterine columnar epithelium thickened, and the mitotic figures in the epithelial stroma and myointimal cells augmented. But no such effect in wet weight and morphology of uterus was found in the GNC group. Conclusion： GNC could increase the BMD and biomechanical parameters of the lumbar vertebrae, reduce the serum TO and LDL-C levels, yet produce no adverse reaction in stimulating proliferation and hypertrophy of uterus.

Effect of Compound Recipe Gengniankang (更年康) on Senile Sexual Hormone and Expression of Estrogen Receptor in Bone of Climacteric Female Rats

Objective： To compare the therapeutic effect of Compound Recipe Gengniankang （更年康, GNK） with that of hormone replacement treatment （HRT） on climacteric female rats with osteoporosis, and to investigate the roles of estrogen and estrogen receptors in the mechanism of osteoporosis. Methods： Climacteric female rats with osteoporosis were chosen and divided into three groups （GNK group, HRT group and control group）. Apoptosis of ovarian granulose cells was measured by terminal-deoxynucleotidyl transferae mediated nick end labeling （TUNEL） assay. Serum level of estrdiol （E2）, follicle stimulating hormone （FSH）, luteinizing hormone （LH） were determined by the method of radioimmunoassay （RIA）. Reverse transcriptase polymerase chain reaction （RT-PCT） technology was used to evaluate the expression of estrogen receptor （ER） in bone. Bone mineral density （BMD） was measured by double energy X-ray absorption （DEXA）. Results： In the climacteric rats, BMD, serum E2, ER mRNA expression in bone decreased remarkably, and serum FSH, LH and apoptosis of ovarian granulose cells increased obviously. After treating with GNK, all the indexes were reversed except serum E2. The increase of E2 was not significant. Conclusion： GNK is effective on climacteric osteoporosis female rats. Its role is performed not by increasing serum E2 but by enhancing ER in the bone and inhibiting apoptosis of ovarian granulose cells. GNK can deter further exhaustion of ovarian function.

Effect of Bushen Yiqi Hexue Recipe (补肾益气和血方) on Pinopodes Expression on Endometrial Surface in Embryo Implantation Dysfunctional Mice

Objective: To explore the therapeutic mechanism of Bushen Yiqi Hexue recipe (补肾益气血方, BYHR), and observe its effect on pinopodes expression on endometrial surface in mifepristone induced embryo implantation dysfunctional mice. Methods: Pregnant Kunming mice were randomly divided into the normal group, the control group and the treated group. Mice in the treated group were administered with BYHR, and those in the normal group and the control group were administered with normal saline, starting from the very first day of pregnancy (Pd1). On the Pd4, mifepristone (RU486) was subcutaneously injected into mice in the control group and the treated group. Pinopodes expression on endometrial surface at 09:30-10:00 pm on Pd4 (regarded as time point 1, T1) and at 09:30-10:00 am on Pd5 (as time point 2, T2) was determined by scanning electron microscopy. Pregnancy rate and average implanted embryos were observed on Pd7. Results: Pregnancy rate and average implanted embryos in the control group were obviously lower than those in the normal group (all P<0.01), while the two parameters were significantly higher in the treated group (P<0.05, P<0.01, respectively) than those in the control group. In the normal group, abundant developing pinopodes were distributed over the whole endometrial surface in T1, and they were altered to a great deal of fully developed pinopodes in T2. But in the control group, only a few pinopodes were expressed locally on endometrial surface in T1, showing a nonsynchronous figure of development, and they completely disappeared in T2. In the treated group, a lot of developing pinopodes were expressed like those in the normal group but somewhat lagged behind, whereas many fully developed pinopodes were expressed in T2. Conclusion: The decrease of the pinopodes in T1 and the cleaning up of them in T2 is possibly the partial mechanism of mifepristone in inducing embryo implantation dysfunction. It is indicated that BYHR could improve pinopodes expression on endometrial surface, eventually better the uterine receptivity and improve the embryo implantation.

Antitumor and synergistic effect of Chinese medicine “Bushen huayu jiedu recipe” and chemotherapy on transplanted animal hepatocarcinoma

AIM： To investigate the antitumor and synergistic effect of Chinese medicine “Bushen huayu jiedu recipe” （recipe for invigorating the kidney, removing blood stasis and toxic substances） and chemotherapy on mice hepatocarcinoma. METHODS： Bushen huayu jiedu recipe （BSHYJDR） consisting of Chinese Cassia Bark, Psoralea, Zedoary, Rhubarb, etc. is equal to 1.5 g/mL liquid of originated herbs after being decoded, filtered, and concentrated. Kunming mice, weighing 18-22 g, were injected with 0.2 mL ascitic hepatocarcinoma H22 containing 1 × 10^7 cells/mL into armpit of the right forelimb of mice. After 24 h, the mice were weighed and randomly divided into tumor-bearing model control group, cisplatin （DDP） group, BSHYJDR high dosage group, low dosage BSHYJDR group, DDP combined with high and low dosage BSHYJDR group, 10 mice in each group. DDP group received injection intraperitoneally （ip） at the dosage of 1 mg/kg （equal to 1/10 LD50）, once a day for 4 d. High and low dosage BSHYJDR groups received intragastric BSHYJDR at the dosages of 26.6 and 13.3 g/kg （20 and 10 times each of clinical adult dosage） respectively, while tumor-bearing model group received the equal volume of distilled water once a day for 10 d. On the 11^th d, the mice were weighed and killed, then the tumor was dissected and weighed, the repression rate （RR） was calculated according to the mean weight of tumor （MWT）. RESULTS： Compared to the model group （MWT： 1.30±0.73）, DDP group （MWT： 0.41±0.09, RR： 68.46%） had a significant difference in the inhibition of hepatocarcinoma H22 （P〈0.01）. High dosage BSHYJDR group （MWT： 0.69±0.29, RR： 46.92%） also had a significant difference in inhibition （P〈0.05）, while no difference was found in low dosage BSHYJDR group （MVVT： 0.85±0.34, RR： 34.62%） （P〉0.05）. When DDP was combined with high dosage BSHYJDR （MWT： 0.29±0.17, RR： 77.69%） and low dosage BSHYJDR （MWT： 0.38±0.21, RR： 70.77%） respectively, we could see improvement of the inhibition effect of DDP on transplanted hepatocarcinoma H22. DDP combined with high dosage BSHYJDR had a significant difference （P〈0.001） compared to DDP, while DDP combined with low dosage BSHYJDR only had a little improvement that is not remarkable. CONCLUSION： Chinese medicine BSHYJDR in combination with chemotherapy can inhibit transplanted hepatocarcinorna in mice.

Clinical Study on Zhuyu Tongfu (逐瘀通腑) Serial Recipe Combined with Acupuncture and Massotherapy in Treating Hypertensive Cerebral Hemorrhage

Objective. To observe the clinical efficacy and mechanism of Zhuyu Tongfu （逐瘀通腑, ZYTF） Serial Recipe combined with acupuncture and massotherapy in treating hypertensive cerebral hemorrhage （HCH）. Methods： One hundred and eighteen patients with hypertensive cerebral hemorrhage, on the basis of conventional Western medicine treatment, were randomly divided into ZYTF combined with acupuncture and massotherapy group （treated group） and simple Western medicine group （control group） ; the clinical efficacy, neurofunction deficit scoring （NDS） alterations and hematoma absorption rate of both groups were observed, and also the plasma superoxide dismutase （SOD） activity, plasma lipid peroxidase （LPO） content, erythrocyte glutathion peroxidase （GSH-Px） activity, hematocrit （Ht） and the whole blood viscosity （Va） change were also observed. Results： In the treated group, the clinical efficacy, NDS improvement and hematoma absorption rate were superior to that of the control group; comparison between the two groups after treatment showed that plasma SOD activity and GSH-Px activity got more elevated and plasma LPO content, Ht and Va more lowered in the the treated group than those in the control group. Cenclusien： ZYTF combined with acupuncture and massotherapy has better effect, its therapeutic mechanism was possibly correlated to the elevation of plasma SOD activity, GSH-Px activity and lowering of plasma LPO content, Ht and Va.

Mechanism of Jianpi Qingchang Huashi Recipe in treating ulcerative colitis:A study based on network pharmacology and molecular docking

BACKGROUND Ulcerative colitis(UC)is a refractory intestinal disease with alternating onset and remission and a long disease course,which seriously affects the health and quality of life of patients.The goal of treatment is to control clinical symptoms,induce and maintain remission,promote mucosal healing,and reduce recurrence.Clinical trials have shown unsatisfactory clinical response rates.As a supplementary alternative medicine,traditional Chinese medicine has a rich history and has shown good results in the treatment of UC.Because of the quality of herbal medicine and other factors,the curative effect of traditional Chinese medicine is not stable enough.The mechanism underlying the effect of Jianpi Qingchang Huashi Recipe(JPQCHSR)on inducing UC mucosal healing is not clear.AIM To investigate the potential mechanism of JPQCHSR for the treatment of UC based on network pharmacology and molecular docking.METHODS Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform was used to extract the active components and action targets of JPQCHSR,and the target names were standardized and corrected through UniProt database.The related targets of UC were obtained through GeneCards database,and the intersection targets of drugs and diseases were screened by jvenn online analysis tool.The visual regulatory network of"Traditional Chinese medicine-active components-target-disease"was constructed using Cytoscape software,the protein interaction network was constructed using STRING database,and enrichment analysis of gene ontology and Kyoto Encyclopedia of Genes and Genomes pathways was conducted through R software.At last,the active components were docked with the core target through SYBYL-X 2.1.1 software.RESULTS Through database analysis,a total of 181 active components,302 targets and 205 therapeutic targets were obtained for JPQCHSR.The key compounds include quercetin,luteolin,kaempferol,etc.The core targets involved STAT3,AKT1,TP53,MAPK1,MAPK3,JUN,TNF,etc.A total of 2861 items were obtained by GO enrichment analysis,and 171 items were obtained by KEGG(Kyoto Encyclopedia of Genes and Genomes)pathway enrichment analysis.The results of molecular docking showed that the key active components in JPQCHSR had certain affinity with the core target.CONCLUSION The treatment of UC with JPQCHSR is a complex process of multi-component,multi-target and multi-pathway regulation.The mechanism of this Recipe in the treatment of UC can be predicted through network pharmacology and molecular docking,so as to provide theoretical reference for it to better play its therapeutic role.

Effect of Bushen Yiqi Huoxue Recipe on Placental Vasculature in Pregnant Rats with Fetal Growth Restriction Induced by Passive Smoking

Interactions of vascular endothelial growth factor （VEGF） with receptors VEGFR1/Fltl and VEGFR2/Flk1, and those of angiopoietins （Ang-1, Ang-2） with receptor Tie2 play important roles in placental angiogenesis. This study investigated vascular morphology and expression of these angiogenic factors in rat placenta on the day 15, 18, 21 of gestation （D 15, D 18 and D21）. The rats were randomly assigned into 3 groups： normal group, model group [fetal growth restriction （FGR） model], and Bushen Tqi Huoxue （BYHR） recipe treatment group （BYHR group, the pregnant rats with FGR were treated with BYHR recipe）. Morphological analysis indicated that during initial villous formation, fetal nucle- ated erythrocytes （FNEs） appeared in maternal blood sinus （MBS）. Subsequently, FNEs were sur- rounded by endothelial cells to form fetal capillary （FC） and then by trophoblast cells to form villi. As pregnancy proceeded, FC density increased progressively with increasing endothelial identification staining （EIS） in normal and BYHR groups. Whereas, villous formation was suppressed, normal in- crease in FC density was impaired and EIS was weakened in model group. Quantitative PCR analysis showed that VEGF and Flkl mRNA increased over gestation in all groups, indicating that VEGF might play a pivotal role in FC growth during late gestation. VEGF mRNA was increased on D15, while de- creased on D21 in model group as compared with normal group and BYHR group. Immunohistochemi- cally, Ang-2 protein was highly expressed in FNEs, gradually disappeared as villi matured, and decreased over gestation in all groups, indicating that Ang-2 might play a pivotal role in villous formation, which was further supported by decreased Ang-2 mRNA and protein expression in model group on D 15. Ang-1 mRNA, Tie2 mRNA and Ang-1/Ang-2 ratio increased from D15 to D18 in all groups as placenta matured. Ang-1 mRNA, Tie2 mRNA and Ang-1/Ang-2 ratio were decreased on D18 in model group as compared with normal and BYHR groups, indicating delayed maturity of FGR placenta. Alterations in angiogenic factors may result in altered placental vasculature and cause placental insufficiency. BYHR recipe could balance the angiogenic factors to promote the formation and maturation of FGR placental vasculature.

Influence of tonifying kidney recipe on advanced glycation endproducts and lipid peroxidation in ovariectomized rats

BACKGROUND： Previous studies have demonstrated that reduced estrogen levels may accelerate the formation of advanced glycation endproducts （AGE） in brain tissue, raise the concentration of lipid peroxidation products in vivo, and speed up deterioration of learning and memory. A tonifying kidney recipe is hypothesized to improve the ability of learning and memory in ovariectomized rats by downregulating AGE and lipid peroxidation products. OBJECTIVE： To simulate a postmenopausal state, bilateral ovariectomy （OVX） was performed in rats, and the effects of tonifying kidney recipe （TKR） on AGE and lipid peroxidation in the rat cerebral cortex, hippocampus, and blood serum levels was measured. In addition, the effects on learning and memory were evaluated, and the effect of AGE -specific inhibitor aminoguanidine （AG） was compared with TKR. DESIGN, TIME AND SETTING： A randomized, in vivo, control experiment was performed at the scientific research center （Provincial Key Laboratory） in the Fourth Hospital of Hebei Medical University （Shijiazhuang, Hebei Province, China） from May 2005 to January 2007. MATERIALS： Forty healthy, adult, female, Sprague Dawley rats were used for this study. TKR was composed of prepared rehmannia rhizome, epimedium herb, desert-living cistanche, and Szechwan lovage rhizome, which were provided by Shijiazhuang Medical Materials Company （China）. A TKR extraction was prepared for further use. AG was provided by Sigma （USA）. Forty rats were randomly divided into four groups： sham, OVX, AG, and TKR, with 10 rats in each group. METHODS： The rat ovaries were resected in the OVX, AG, and TKR groups, whereas the same volume of fat was resected in the sham group. At four weeks after OVX, the AG group received 1% AG water solution by lavage; the TKR group was administrated by lavage once per day at a dose of 6.3 g （crude drug）/kg; OVX and sham groups received equal volumes of tap water. MAIN OUTCOME MEASURES： Learning and memory behavior of rats was tested in a Y-electric maze 16 weeks after the OVX procedure. The contents of advanced glycation endproducts in the rat cerebral cortex, hippocampus, serum, and urine were detected by competitive ELISA and spectrofluorophotometer. The contents of lipid peroxidation in rat serum, cerebral cortex, and hippocampus were assayed using a biochemical method. RESULTS： Compared with the sham group, serum content of advanced glycation endproducts in the OVX group was significantly increased, and lipid peroxidation content increased in cerebral cortex, hippocampus, and serum （t = 3.04-4.22, P 〈 0.05 0.01）. Both AG and TKR decreased the amount of AGE in cerebral cortex and serum （t = 2.53, 3.64, P 〈 0.05, 0.01）, increased AGE urine content （t = 3.25 4.87, P 〈 0.01）, and decreased lipid peroxidation content in cerebral cortex, hippocampus, and serum （t = 2.80 3.70, P 〈 0.05 0.01）. In comparison to the OVX and sham groups, the correct escape rate in the Y-electric maze was significantly increased （t = 3.46, 3.28, P 〈 0.01）, and escape latency was significantly decreased （t=3.12, 2.48 P 〈 0.05） in the AG and TKR groups, which indicated that both AG and TKR improved learning and memory The OVX group had a significantly lower correct escape compared with the sham group （t = 4.21, P 〈 0.01 ）.CONCLUSION： The tonifying kidney recipe decreased deposition of advanced glycation endproducts and lipid peroxidation in ovariectomized rats, and concomitantly improved learning and memory. The effect of TKR was equal to that of AG.

Effect of Bushenantai Recipe on the Expression of Endometrial LIF in Mice with Embryonic Implantation Dysfunction

In order to observe the effect of Bushenantai recipe on the expression of endometrial leukemia-inhibitory factor （LIF） in mice with embryonic implantation dysfunction （EID）, 120 Kunming mice post coition were randomized into three groups： normal control group, model group and traditional Chinese medicine group （TCM group） （n=40 in each group）. Uterus was collected on the pregnancy day （Pd） 4, 5, 6 after an intravenous injection of Evan＇s blue. The endometrium was dyed by Evan＇s blue and the mean points of response were observed on Pd 5. The expression of LIF mRNA and protein was detected by RT-PCR and immunohistochemistry respectively and analyzed statistically by image system. The results showed that the number of implantation sites in model group was remarkably less than in normal control group and TCM group. There was no significant difference between normal control group and TCM group. The expression of LIF mRNA and protein in model group was delayed. Bushenantai recipe could increase the expression of LIF mRNA and protein in endometria of mice with EID. It was suggested that Bushenantai recipe could improve embryo implantation of mice with EID by promoting the endometrial LIF expression and endometrial decidualization.

Effect of Yiqitongyanghuatan Recipe on Experimental Atherosclerosis in Rabbits

In order to investigate the roles of Yiqitongyanghuatan (YQTYHT) recipe in reducing the levels of serum cholesterol and plasma lipid peroxidation(LPO), platelet aggregation function (PAgF) and platelet adhesion function (PAdF), the area of atherosclerotic plague coverage in aorta and the thickness of plague, 32 male Japanese white rabbits were divided into 4 groups. The results showed that the YQTYHT recipe could significantly lower the levels of serum cholesterol and tryglyceride, plasma LPO, and PAgF and PadF. The area of atherosclerotic plague coverage in aorta and the thickness of plague in the YQTYHT fed rabbits were decreased as compared with that in the high cholesterol fed rabbits. The above roles might contribute to the main mechanism of YQTYHT against atherosclerosis.

Influence of Shenqing Recipe on Morphology and Quantity of Colonic Interstitial Cells of Cajal in Trinitrobenzene Sulfonic Acid Induced Rat Colitis

Objective To observe the influence of Shenqing Recipe (SQR), a kind of Traditional Chinese Medicine, on the morphology and quantity of colonic interstitial ceils of Cajal (ICC) in trinitrobenzene sulfonic acid (TNBS)-induced rat colitis, and to investigate the possible mechanism of SQR in regulating intestinal dynamics. Methods Sixty rats were randomly divided into normal control, model Ⅰ , model Ⅱ, mesalazine, and high-dose, and low-dose SQR groups with 10 rats in each group. TNBS (10 mg) dissolved in 50% ethanol was instilled into the lumen of the rat colon of the latter five groups to induce colitis. On the 4th day after administration of TNBS, each treatment group was administered one of the following formulations by enteroclysis gavage once a day for 7 days: 600 mg·kg^-1·d^-1 mesalazine, 2.4g^-1·d^-1 SQR, and 1.2g^-1·d^-1 SQR. Model Ⅱ rats received normal saline solution. After 7 days colonic samples were collected. While the colonic samples of model I group were collected on the 3rd day after TNBS administered. Ultrastructure of ICC in the damaged colonic tissues was observed with transmission electron microscope. Expression of c-kit protein in colonic tissue was determined by immunohistochemical staining and Western blot. Results The ultrastructure of colonic ICC in the rat model of TNBS-induced colitis showed a severe injury, and administration of SQR or mesalazine reduced the severity of injury. Similarly, the expression of c-kit protein of TNBS-induced colitis rat model was significantly decreased compared with the normal control group (P〈0.05). Treatment with SQR or mesalazine significantly increased the expression of c-kit protein compared with the administration of control formulations (P〈0.05), especially the high-dose SQR group. Conclusion SQR could alleviate and repair the injured ICC, and improve its quantity, which might be involved in regulating intestinal motility.