Regulation of non-classical immune parameters in immune thrombocytopenic purpura mice by a spleen-invigorating,qi-replenishing and blood-containing formula

Objective:This study investigated the regulatory effect of non-classical immune parameters on immune thrombocytopenic purpura(ITP)mice by a spleen-invigorating,qi-replenishing and blood-containing formula(SQBF).Method:A total of 80 BALB/c mice were randomly divided into four equal groups(20 mice each):control group,model group,prednisone group and spleen-invigorating,qi-replenishing and blood-containing(SQBF)group.Mice in the model group,prednisone group,and SQBF group were administered anti-platelet serum to induce ITP.The dynamic variations of platelet counts in ITP mice were measured with an automatic blood analyzer before modeling and 48 h,and 8,12 and 15 days following APS injection.Levels of b-endorphin(b-EP),vasoactive intestinal peptide(VIP)and salivary IgA(SIgA)were detected by enzyme-linked immunosorbent assay(ELISA)on 15th day of experiment.Results:SQBF enhanced peripheral blood platelet counts in ITP mice similar to that of prednisone,and both groups showed a statistically significant response compared with the model group(P<.01).The SQBF significantly decreased b-EP levels compared with the model and prednisone intervention groups(P<.05),significantly increased the levels of VIP and SIgA in ITP mice compared with the model group(P<.05)and had significant protective effects on the thymus of ITP mice compared with the model group(P<.01).Conclusions:The SQBF had a similar effect to prednisone with regards to enhancing peripheral blood platelet counts in ITP mice.Furthermore,it decreased b-EP levels and increased VIP and SIgA,and protected the thymus.This shows that,on base of the brain-gut axis functions,some non-classical immune vascular active factors or neurotransmitters are also involved in immune responses,and also have relationship with the onset of ITP and bleeding and/or hemostasis.It needs further study to determine whether a change in these active factors is related to immediate hemostasis.

Effect of Yang-Warming and Kidney-Tonifying Prescription on Expression of Osteogenic and Angiogenic Factors and H-Type Vascular Markers in Steroid-Induced Avascular Necrosis of Femoral Head in Rabbits

Objective: To investigate the effect of Yang-warming and Kidney-tonifying Prescription (YKP) on the treatment of steroid-induced avascular necrosis of the femoral head (SANFH) in rabbits. And to further explore whether its therapeutic mechanism is related to the expression of HIF-1α and VEGF (angiogenic factors), BMP2 and Osterix (osteogenic factor), CD31 (type H vascular marker) and MMP13 (bone destruction-related factor). Methods: Twenty-seven healthy male New Zealand white rabbits were divided into a normal group, model group, traditional Chinese medcine (TCM) group (clinical equivalent dose group of YKP), miR-130a inhibitor group and TCM + inhibitor group. The SANFH model was established by combining horse serum with methylprednisolone. After the model is successfully established, TCM group was given 6.44 g/kg·d YKP by gavage, and the miR-130a gene inhibitor group was intraperitoneally injected with 25 mg/kg miR-130a inhibitor, locked nucleic acid (LNA)-anti-miR-130a. TCM + inhibitor group was treated with YKP intragastrically and miR-130a inhibitor intraperitoneally. The rabbits in the normal group and the model group were intragastrically administered with normal saline 10 ml/d. Once a day for 4 weeks. The avascular necrosis was detected by HE staining. The contents of HIF-1α, VEGF, BMP2 and Osterix in rabbit tissues were detected by qRT-PCR kit, and the expression of CD31 and MMP13 was detected by immunofluorescence staining. Results: In the normal group, the surface of the cartilage layer of the femoral head was smooth, the bone trabeculae were intact and densely arranged, the cells of each layer were neatly arranged, the morphology of the bone cells, the chondrocytes and the adipocytes were normal. In the model group, cartilage surfaces of the femoral head showed exfoliative cracks. The bone trabecular structure was loose and incomplete, chondrocytes, osteoblasts and bone marrow cells were significantly reduced, and the number of empty bone traps was significantly increased. In the TCM-treated group, more chondrocytes, thicker cartilage layer, and more regular bone trabeculae were detected as compared to model rabbits. In contrast, the cartilage layer was thinner, the destruction and fracture of bone trabeculae was more serious, chondrocytes and osteocytes were decreased as compared to model group. The expression of HIF-1α, VEGF, BMP2, and Osterix in the model group decreased significantly as compared to the normal group (P Conclusion: YKP can regulate the expression of angiogenic-related factors (VEGF and HIF-α), osteogenic-related factors (BMP2 and Osterix), and H-type vascular marker CD31, resulting in increased expressions of VEGF, HIF-α, BMP2, and Osterix, which promote intra-femoral head revascularization. Meanwhile, YKP decreased the expression of bone-destruction-related factor MMP13, thus enhancing the ability of bone tissue to repair itself. Regulation of these molecules’ expression may be one of the mechanisms of YKP in the treatment of hormonal femoral head necrosis.

Meta-analysis on the treatment of diabetic nephropathy by spleeninvigorating,kidney-tonifying and blood-activating method combined with western medicine

Objective:This article uses a meta-analysis system to evaluate the clinical efficacy of the method of spleen-invigorating,kidney-tonifying and blood-activating combined with western medicine in the treatment of diabetic nephropathy,in order to provide evidence-based medicine for the clinical treatment of this disease.Methods:Computer search(January 2009 to June 2020)published in CNKI,Wanfang database,and Weipu Chinese Science and Technology Journal Database with simple western medicine conventional treatment(control group)and on the basis of control group Randomized Controlled Trial(RCT)of the treatment of diabetic nephropathy with spleen-invigorating,kidney-tonifying and blood-activating(experimental group),and manual retrieval of relevant literature that meets the inclusion criteria.Two researchers independently carried out literature screening,and used the improved Jadad rating scale for quality evaluation,and finally used RevMan5.3 statistical software for meta-analysis.Results:A total of 15 randomized controlled trials were included,with a total of 1218 patients,including 621 in the experimental group and 597 in the control group.Meta-analysis showed that the experimental group was improving clinical efficacy(OR=4.35,95%CI[3.12,6.08],P<0.00001),FPG(MD=-0.49,95%CI[-0.89,-0.09],P=0.02),24h urine protein quantification(MD=-0.54,95%CI[-0.90,-0.18],P=0.003),mALB(MD=-15.95,95%CI[-18.12,-13.79],P<0.0001),BUN(MD=-1.79,95%CI[-2.99,-0.59],P=0.004),SCr(MD=-47.63,95%CI[-83.43,-11.83],P=0.009),TC(MD=-1.02,95%CI[-1.42,-0.62],P<0.00001),etc.may be better than the control group;The test group is improving TG(MD=-1.02,95%CI[-1.42,-0.62],P<0.00001)there is no significant difference compared with the control group.Conclusion:The method of spleen-invigorating,kidney-tonifying and blood-activating combined with conventional western medicine treatment of diabetic nephropathy may be superior to conventional western medicine treatment in terms of improving clinical efficacy,FPG,24h urine protein quantification,mALB,BUN,SCr,TC,etc.However,due to the small number and low quality of the literature included in this study,more in-depth research needs to further expand the sample size and include higher quality RCTs.

Review for treatment and signaling pathway regulation of kidney-tonifying traditional Chinese medicine on osteoporosis

The treatment and signaling pathway regulation effects of kidney-tonifying traditional Chinese medicine on osteoporosis have been widely studied,but without a systematic summary currently.This review comprehensively collected and analyzed the traditional Chinese medicine on the treatment and signaling pathway regulation of osteoporosis in recent ten years,such as Epimedium,Drynariae Rhizoma,Cnidium,Eucommia,Psoralen and Dipsacus.Based on the existing findings,we concluded the following conclusions:(1)kidney-tonifying traditional Chinese medicine treats osteoporosis mainly through BMP-Smads,Wnt/β-catenin,MAPK,PI3K/AKT signaling pathway to promote osteoblast bone formation and through OPG/RANKL/RANK,estrogen,CTSK signaling pathway to inhibit osteoclasts of bone resorption.(1)Epimedium,Drynariae Rhizoma,Cnidium and Psoralen up-regulate the key proteins and genes of BMP-Smads and Wnt/β-catenin signaling pathways to promote bone formation.(2)Epimedium,Drynariae Rhizoma,Cnidium,Eucommia,Psoralen,Dipsacusinhibit the bone resorption by mediating the OPG/RANKL/RANK signaling pathway.(2)Kidney-tonifying traditional Chinese medicine prevent and treat osteoporosis through a variety of ways:Icariin,Naringin,Osthol,Psoralen can regulate BMP-Smads,Wnt/β-catenin signaling pathway to promote bone formation,but also activate OPG/RANKL/RANK,CTSK and other signaling pathway to inhibit bone resorption.(3)The crosstalk of the signaling pathways and the animal experiments of the traditional Chinese medicine on the prevention and treatment of osteoporosis as well as their multi-target mechanism and comprehensive regulation need further clarification.

Protective effects of kidney-tonifying Chinese herbal preparation on substantia nigra neurons in a mouse model of Parkinson's disease

The Chinese herbs Herba Epimedii, Fructus Ligustri Lucidi and Rhizoma Polygonati were injected into Parkinson＇s disease mice established via intraperitoneal injection of 1-methyl-4-phenyl-1, 2, 3, 6-tetrahydropyridine hydrochloride. The selective monoamine oxidase B inhibitor selegiline was used as a positive control drug. After successive administration for 4 weeks, Herba Epimedii could downregulate the expression of caspase-3 and increase the brain-derived neurotrophic factor level, as well as increase tyrosine hydroxylase activity in the substantia nigra of Parkinson＇s disease mouse models. Rhizoma Polygonaticould downregulate the expression of caspase-3 and FasL, and increase neural growth factor and brain-derived neurotrophic factor levels. Fructus Ligustn Lucidi could downregulate caspase-3 expression. Rhizoma Polygonati and Fructus Ligustn Lucidi did not produce obvious effects on tyrosine hydroxylase activity. Herba Epimedii and Fructus Ligustri Lucidi yielded similar effects on apoptosis-promoting factors to those elicited by selegiline. Herba Epimedii and Rhizoma Polygonati significantly increased the levels of neurotrophic factors compared with selegiline. Herba Epimedii significantly increased tyrosine hydroxylase activity compared with selegiline. It is indicated that the kidney-tonifying Chinese herbal preparation can downregulate the expression of apoptosis-promoting factors, increase neurotrophic factors levels in the substantia nigra and striatum, as well as increase tyrosine hydroxylase activity in the substantia nigra of Parkinson＇s disease mouse models, thereby exerting a stronger or similar neuroprotective effects compared with selegiline.

Meta analysis of infertility due to polycystic ovarian syndrome treated by kidney-tonifying and blood-activating therapy

Objective:To explore the Meta-analysis of polycystic ovary syndrome-induced infertility by tonifying kidney and activating blood circulation.Methods:Chinese keywords or subject words such as"kidney-tonifying and Blood-Activating Therapy","polycystic ovary syndrome","infertility"were searched in CNKI,Wanfang and Weipu databases,and English keywords such as"Kidney-tonifying and Blood-Activating Therapy","Stein-Leventhal syndrome","Infertility"were searched in Pubmed database for a period of 2.From 2000 to 2019,Meta-analysis was carried out with Review Manager 5.0.Results:Thirteen randomized controlled literatures were screened out,and the control group was treated with routine western medicine and the treatment group was treated with Kidney-tonifying and Blood-activating therapy.All of them included literatures mentioned grouping method,but no blind method was mentioned,and the literature quality was generally low.Meta-analysis of the effective rate showed that Z=3.69(P=0.0002<0.001),95%CI(1.77,6.42),indicating that the effective rate in the treatment group was higher than that in the control group.Meta-analysis of the cycle ovulation rate showed that Z=2.19(P=0.03<0.05),95%CI(1.08,4.35),indicating that the cycle ovulation rate in the treatment group was higher than that in the control group.Meta-analysis of pregnancy rate showed that:Z=7.07(P<0.00001),95%CI(2.27,4.24),indicating that the pregnancy rate in the treatment group was higher than that in the control group;bias analysis found that the literature was distributed on both sides of the dotted line,most of which were biased.Meta-analysis of the incidence of LUFS showed that Z=3.25(P=0.001<0.05),95%CI(0.02,0.38),indicating that the incidence of LUFS in the treatment group was lower than that in the control group.Meta-analysis of serum E2 level after treatment showed that Z=4.41(P<0.0001),95%CI(11.01,28.64),indicating that the serum E2 level in treatment group was higher than that in control group after treatment.Conclusion:Meta-analysis shows that the method of tonifying kidney and activating blood circulation has a significant effect on infertility caused by polycystic ovary syndrome.It can improve the level of sex hormones,improve the efficiency,cycle ovulation rate and pregnancy rate,and reduce the incidence of LUFS.However,the quality of the included literature is not high,which needs to be further included in high-quality literature to confirm.

Effects of kidney-tonifying and blood circulation-promoting recipes and platelet-rich plasma on the activity and expression of type I collagen

Objective:To observe the effects of kidney-tonifying and blood circulation-promoting recipes and platelet-rich plasma(PRP)on the activity and expression of type I collagen in co-culture system.Methods:The subcutaneous adipose tissue and degenerative nucleus pulposus tissue of patients with nucleus pulposus removal under lumbar intervertebral disc degeneration were isolated and cultured by multiple enzyme digestion methods to establish a co-culture system of ADSCs/NPCs.Patients with lumbar intervertebral disc degeneration were enrolled for 2 weeks.The venous blood was taken 50mL before the first dose and one hour after the last dose.Platelet-rich plasma was prepared by secondary centrifugation and cultured with different plasma.The experiment was divided into five groups:platelet-rich plasma group(PRP),platelet-poor plasma group(PPP),drug-rich platelet-rich plasma group(M-PRP)and drug-poor platelet plasma group(M-PPP),fetal bovine serum group(FBS).The morphological changes of the cells in each group were observed under an inverted microscope.The changes of type I collagen activity were detected by immunofluorescence assay.The expression of CCOL1A1 gene was detected by RT-PCR two weeks after the intervention.Results:In this study,the results of kidney-tonifying and blood circulation-promoting recipes and PRP group intervention were shown.The results of the group intervention showed that the M-PRP group and the PRP group had clearer cell contours and better refractive index than the other groups,and there was no significant difference between the two groups.The results of immunofluorescence staining showed that the number of cells in PRP group and M-PRP group was higher,but the staining intensity of single cells was significantly lower than that in FBS group.RT-PCR results showed that the expression of COL1A1 mRNA in PPP group,PRP group,M-PRP group and M-PPP group was significantly down-regulated(P<0.05),respectively:0.343±0.040,0.294±0.018,0.310±0.022,0.430±0.020.Conclusions:Kidney-tonifying and blood circulation-promoting recipes combined with platelet-rich plasma can decrease the activity and exhibit a tendency to inhibit the expression of type I collagen.

Effects of Kidney-tonifying and Blood Circulation-promoting Recipes and PRP on the Activity and Expression of Type I Collagen

Objective:To observe the effects of kidney-tonifying and blood circulation-promoting recipes and platelet-rich plasma (PRP) on the activity and expression of type I collagen in co-culture system.Methods:The subcutaneous adipose tissue and degenerative nucleus pulposus tissue of patients with nucleus pulposus removal under lumbar intervertebral disc degeneration were isolated and cultured by multiple enzyme digestion methods to establish a co-culture system of ADSCs/NPCs. Patients with lumbar intervertebral disc degeneration were enrolled for 2 weeks.The venous blood was taken 50mL before the first dose and one hour after the last dose. Platelet-rich plasma was prepared by secondary centrifugation and cultured with different plasma. The experiment was divided into five groups: platelet-rich plasma group (PRP), platelet-poor plasma group (PPP), drug-rich platelet-rich plasma group (M-PRP) and drug-poor platelet plasma group (M-PPP), fetal bovine serum group (FBS). The morphological changes of the cells in each group were observed under an inverted microscope. The changes of type I collagen activity were detected by immunofluorescence assay. The expression of CCOL1A1 gene was detected by RT-PCR two weeks after the intervention.Results:In this study, the results of kidney-tonifying and blood circulation-promoting recipes and PRP group intervention were shown. The results of the group intervention showed that the M-PRP group and the PRP group had clearer cell contours and better refractive index than the other groups, and there was no significant difference between the two groups. The results of immunofluorescence staining showed that the number of cells in PRP group and M-PRP group was higher, but the staining intensity of single cells was significantly lower than that in FBS group. RT-PCR results showed that the expression of COL1A1 mRNA in PPP group, PRP group, M-PRP group and M-PPP group was significantly down-regulated (P<0.05), respectively: 0.343±0.040, 0.294±0.018, 0.310±0.022, 0.430±0.020.Conclusion:Kidney-tonifying and blood circulation-promoting recipes combined with platelet-rich plasma can decrease the activity and exhibit a tendency to inhibit the expression of type I collagen.

Investigation of hemostatic effect of Spleeninvigorating,Qi-replenishing and Bloodarresting Formula on simvastatin-induced zebrafish hemorrhage model

Blood-arresting Formula on simvastatin-induced zebrafish hemorrhage model compared with that of the Heat-clearing and Blood-cooling Formula.Methods:AB strain wild type zebrafish were treated with 0.5 mM simvastatin for 24 hours to establish the zebrafish hemorrhage model.Under strictly blind experimental conditions,the zebrafish were then treated with experimental agents in different concentration within the maximum non-lethal dose.The intervention effect of the Spleen-invigorating,Qi-replenishing and Blood-arresting Formula was comprehensively assessed by examining the main observational parameters such as the bleeding reduction rate and hemostatic rate while referring to the additional parameters such as blood flow,improvement rate of blood flow,velocity of movement,improvement rate of motion under spleen-qi deficiency.Results:Hemostatic effect:The bleeding rate and hemostatic rate in the zebrafish were respectively,30%,15%,45%,40%and 60%,80%,40%,47%when equal concentration of 500 mg/mL,1000 mg/mL were used.This showed that the experimental herbs B1 was superior to B2 in decreasing the bleeding rate and improving hemostatic effect in the zebrafish.Improvement of blood flow:With equal concentration,the experimental herb B1 was superior to B2 in increasing the blood flow and improvement rate of blood flow in hemorrhagic zebrafish.Promotion of the improvement of motion:With equal concentration of 500 mg/mL and 1000 mg/mL,the experimental herb B1 was superior to B2 in promoting the velocity of motion in the zebrafish and promoting the improvement rate of motion.Conclusion:The manifestations of simvastatin-induced zebrafish hemorrhage are basically similar to that of failure of the spleen to control blood in human beings.The Spleeninvigorating,Qi-replenishing and Blood-arresting Formula has good hemostatic effect on simvastatin-induced hemorrhage in the zebrafish.It also boasts the functions of improving blood flow and velocity of motion in the hemorrhagic zebrafish.Such a conclusion provides the experimental basis for the treatment of the syndrome of failure of the spleen to control blood by this formula.

Investigation of Mechanism of Premature Ovarian Failure Regulation by Kidney-tonifying Herbs and Liver-clearing Herbs in Dingjing Decoction

Objective To investigate the mechanisms through which kidney-tonifying herbs（KTHs） and liver-clearing herbs（LCHs） in Dingjing Decoction（DJD） regulate premature ovarian failure（POF）. Methods One hundred and fifty Sprague-Dawley rats were randomly divided into five groups such as control, model, KTHs, LCHs, and DJD groups. POF-related biological molecules were examined. Factor analysis was performed to investigate the regulatory networks and key biomolecules involved in mediating POF after treatment with KTHs and LCHs. Results The master regulatory factors in the reproductive endocrine network associated with KTHs intervention included four molecules in the pituitary-ovarian axis, cortisol（CORT） in the target gland of pituitary-adrenal axis, and some molecules in the hypothalamus. In contrast, the master regulatory factors associated with LCHs intervention included four molecules in the pituitary-ovarian axis and some molecules in the hypothalamus; No biomolecules in the pituitary-adrenal axis were involved in the LCH-mediated mechanisms.Gonadotropin-releasing hormone（Gn RH）, which was identified as a common biological molecule in the hypothalamus, was involved in regulating the reproductive endocrine network in association with KTHs intervention. Conclusion KTHs directly regulates biological molecules in the pituitary-adrenal axis and indirectly regulates those in the pituitary-adrenal axis through the hypothalamus, while the LCHs only exert its effects indirectly. Gn RH is the key biological molecule associated with KTHs intervention.

Effects of Chinese Kidney-Tonifying Herbal Medicine on M-Cholinergic Receptor and Acetylcholinesterase Activity in Dementia Mimetic Mice

Objective: In order to study the effects of Chinese Kidney-tonifying (KT) herbal medicine on the M-cholinergic receptor(M-R) and the acetylcholinesterase (AchE ) activity in the models' cerebral cortex. Methods: Dementia mimetic mice models were formed with aluminum chloride. Results: The Muscarine receptor total receptor concentration (M-Rt) lowered and the AchE increased in the model significantly comparedwith healthy young mice. KT could markedly reduce the increase of AchE and elevate the lowered M-Rt in cerebral cortex of the dementia mimetic mice and improve the memory. Conclusions: KT is effective in preventingthe degeneration of cerebral function and presenile dementia.

Effects of Er'zhi Tiangui Granule (二至天癸颗粒) on Sequential Expressions of Integrin β3 and Its Ligand Osteopontin in Mouse Endometrium during Controlled Ovarian Hyperstimulation

To investigate the effects of ECzhi Tiangui Granule （二至天癸颗粒, ETG） on sequential expressions of integrin β 3 and its ligand osteopontin in the mouse endometrium during controlled ovarian hyperstimulation （COH） and implantation period. Methods： Seventy-five Mature female Kunming mice were randomly divided into 3 groups, a normal control group, a model group, and a treatment group administrated with ETG for 10 days, 25 in each group. After mated with male mice, every 5 mice were sacrified in each group at the 0, 2nd, 4th, 6th, and 8th days to take their endometrium. In-situ hybridization was used to detect the expressions of integrin β 3 and osteopontin in the endometrium. Results： mRNA expressions of integrin β 3 and osteopontin in the endometrium during implantation period showed similar time sequence rules in the treatment group to those in the normal control group; the peak values of them were a little lower in the treatment group than the normal control without significant differences. In the model group, integrin β 3 mRNA expression was higher at the 2nd day, obviously lower at the 4th and 6th days, and insignificantly lower at the 8th day; and osteopontin expression was remarkably lower at the 4th, 6th, and 8th days, compared with the normal control and the treatment groups （P〈0.05, P〈0.01）. Conclusions： COH might influence the sequential expressions of integrin β 3 and its ligand osteopontin, bring forward the integdn β 3 expression peak, impact on the cooperation of integrin β 3 and osteopontin, so as to damage the endometrial receptivity. ETG could regulate the sequential expressions of integrin β 3 and its ligand osteopontin to improve the mouse endometrial receptivity during COH.