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Natto is a soybean product fermented by natto bacteria. It is rich in a variety of amino acids, vitamins, proteins and active enzymes. It has a number of biological activities, such as thrombolysis, prevention of osteoporosis, antibacterial, anticancer, antioxidant and so on. It is widely used in medicine, health-care food, biocatalysis and other fields. Natto is rich in many pharmacological active substances and has significant medicinal research value. This paper summarizes the pharmacological activities and applications of natto in and outside China, so as to provide references for further research and development of natto.
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The aim of this study was to prepare tandem multimeric proteins of BmSPI38, a silkworm protease inhibitor, with better structural homogeneity, higher activity and stronger antifungal ability by protein engineering. The tandem multimeric proteins of BmSPI38 were prepared by prokaryotic expression technology. The effects of tandem multimerization on the structural homogeneity, inhibitory activity and antifungal ability of BmSPI38 were explored by in-gel activity staining of protease inhibitor, protease inhibition assays and fungal growth inhibition experiments. Activity staining showed that the tandem expression based on the peptide flexible linker greatly improved the structural homogeneity of BmSPI38 protein. Protease inhibition experiments showed that the tandem trimerization and tetramerization based on the linker improved the inhibitory ability of BmSPI38 to microbial proteases. Conidial germination assays showed that His6-SPI38L-tetramer had stronger inhibition on conidial germination of Beauveria bassiana than that of His6-SPI38-monomer. Fungal growth inhibition assay showed that the inhibitory ability of BmSPI38 against Saccharomyces cerevisiae and Candida albicans could be enhanced by tandem multimerization. The present study successfully achieved the heterologous active expression of the silkworm protease inhibitor BmSPI38 in Escherichia coli, and confirmed that the structural homogeneity and antifungal ability of BmSPI38 could be enhanced by tandem multimerization. This study provides important theoretical basis and new strategies for cultivating antifungal transgenic silkworm. Moreover, it may promote the exogenous production of BmSPI38 and its application in the medical field.
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Animals , Antifungal Agents/pharmacology , Escherichia coli/metabolism , Proteins/metabolism , Protease Inhibitors/chemistry , Bombyx/chemistry , Saccharomyces cerevisiae/metabolism , Peptide HydrolasesABSTRACT
Objective:To compare the early efficacy of proximal femoral bionic nail (PFBN) and proximal femoral nail anti-rotation (PFNA) in the treatment of intertrochanteric fracture in the elderly.Methods:A retrospective cohort study was conducted to analyze the clinical data of 45 elderly patients with intertrochanteric fracture treated at First Affiliated Hospital of Henan Polytechnic University from March 2020 to December 2021, including 13 males and 32 females; aged 70-96 years [(78.6±7.3)years]. According to the AO classification, there were 10 patients with type 31-A1, 16 type 31-A2 and 19 type 31-A3. There were 20 patients treated with PFBN (PFBN group) and 25 with PFNA (PFNA group). The operation time, intraoperaive blood loss, bone healing time, and time to full weight-bearing were compared between the two groups. The tip-apex distance, distance from tail of lag screw to lateral side of intramedllary nail, proximal femoral length and femur neck-shaft angle were compared between the two groups at day 1 postoperatively and at the last follow-up, and their amount of changes between the two time points were also measured. Harris hip score was evaluated at 3, 6 months postoperatively and at the last follow-up. Complications were observed as well.Results:All patients were followed up for 6-21 months [(14.7±3.6)months]. There were no significant differences in operation time, intraoperative blood loss and bone healing time between the two groups (all P>0.05). The time to full weight-bearing was significantly earlier in PFBN group [(7.9±1.2)weeks] than that in PFNA group [(9.1±0.9)weeks] ( P<0.05). At day 1 postoperatively and at last follow-up, the tip-apex distance was (23.4±1.7)mm and (23.3±1.6)mm in PFBN group ( P>0.05), and was (24.5±2.1)mm and (24.3±2.3)mm in PFNA group ( P>0.05); the distance from tail of lag screw to lateral side of intramedllary nail was (8.1±1.1)mm and (11.8±0.9)mm in PFBN group ( P<0.01), and was (7.7±1.0)mm and (12.6±1.6)mm in PFNA group ( P<0.01); the proximal femoral length was (91.3±1.6)mm and (88.5±2.6)mm in PFBN group ( P<0.01), and was (91.4±2.0)mm and (87.6±2.0)mm in PFNA group ( P<0.01); the femur neck-shaft angle was (127.2±2.9)°and (125.7±3.0)° in PFBN group ( P>0.05), and was (128.5±3.0)° and (127.2±3.2)° in PFNA group ( P>0.05). There were no significant differences in the above indicators between the two groups at day 1 postoperatively and at the last follow-up (all P>0.05). The amount of changes in the tip-apex distance and femur neck-shaft angle were (-0.2±0.1)mm and (-1.6±0.7)° in PFBN group, similar with (-0.2±0.2)mm and (-1.5±1.0)° in PFNA group (all P>0.05). However, the amount of changes in the distance from tail of lag screw to lateral side of intramedllary nail and proximal femoral length were (3.6±1.4)mm and (2.7±1.2)mm in PFBN group, significantly lower than (5.2±1.1)mm and (4.0±1.1)mm in PFNA group (all P<0.01). There were no significant differences in Harris hip score between the two groups at 3, 6 months postoperatively or at the last follow-up (all P>0.05). PFBN group had 1 patient with fracture displacement, 2 with compression screw backup and 2 with thigh pain. PFNA group had 1 patient with fracture displacement and 4 with thigh pain. Conclusion:For elderly patients with intertrochanteric fracture, PFBN provides not only earlier full weight-bearing, but also stronger stability of fracture fixation than PFNA.
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Objective To explore the variation trend of natural killer (NK) cell subsets in the recipients infected with cytomegalovirus (CMV) after renal transplantation. Methods Clinical data of 92 renal transplant recipients were retrospectively analyzed. All recipients were divided into the CMV infection group (n=43), CMV infection recovery group (n=13), stable renal function group (n=15), rejection group (n=11) and other infection group (n=10). In addition, healthy adult volunteers were enrolled in the healthy control group (n=15). The proportion of NK cells in peripheral blood, the expression proportion and the mean fluorescence intensity (MFI) of CD226 and CD16 in NK cells were observed and statistically compared among different groups. Results The proportion of NK cells was 4.9% (2.2%, 11.5%) in the CMV infection group and 3.7% (2.3%, 6.5%) in the CMV infection recovery group, which were significantly lower than those in the other groups (all P < 0.05). The expression proportion of CD226 and CD16 in NK cells in the CMV infection group was significantly lower compared with those in the healthy control group and stable renal function group(all P < 0.05). The expression proportion of CD226 and CD16 in NK cells in the CMV infection recovery group was remarkably higher than those in the CMV infection group (both P < 0.05). The MFI of CD226 and CD16 in the CMV infection group was significantly lower than those in the healthy control group (both P < 0.05). The MFI of CD226 and CD16 in the CMV infection recovery group was significantly higher than those in the CMV infection group (both P < 0.05). Conclusions The expression proportion and MFI of CD226 and CD16 in NK cells are down-regulated in CMV infection period, whereas up-regulated during the CMV infection recovery period, prompting that CD226 and CD16 expressed by NK cells are intimately correlated with the course of CMV infection.
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Objective To evaluate our noval rod-screw construct in anterior single-segmental decompression and fixation for the treatment of Denis type B burst fractures.Methods From February 2007 through May 2010,we treated 28 patients with Denis type B burst fracture.They were aged from 18 to 52 years (average,37.4 years).The fractures were located at T1 1 in 3 cases,at T12 in 10,L1 in 12,and L2 in 3.Of them,7 were complicated with unilateral pedicle fracture and 3 with bilateral pedicle fractures.According to the American Spinal Injury Association (ASIA) scale,the neurological deficits were rated as grade B in 11 cases,as grade C in 9,and grade B in 8.All the patients received anterior single-segmental decompression,titanium mesh and autogenous bone graft,and fixation with our noval rod-screw construct.The therapeutic efficacy was evaluated in terms of visual analogue scale (VAS),cobb angle,and spinal canal encroachment.Results The mean follow-up time was 26 months (range,from 17 to 33 months).The average length of surgery was 144 minutes(range,from 90 to 176 min);the mean blood loss was 580 mL (range,from 300 to 1 100 mL).The mean VAS score,cobb angle and spinal canal encroachment were improved from 8.2 ± 1.5,17.4° ± 4.3° and 53％ ± 16％ preoperatively to 2.7 ± 2.4,9.4° ± 5.8° and 4％ ±2％ at the final follow-up,respectively(P ＜ 0.05).None of the patients exhibited neurological deterioration.Conclusion Our noval rod-screw construct is a safe and effective device that can be used in the anterior single-segmental decompression and fixation for the treatment of Denis type B burst fractures.
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BACKGROUND:Exogenous basic fibroblast growth factor (bFGF) plays an important role in the ligament tissue healing process, and the use of transgenic methods to transfect exogenous genes into cells can promote the secretion of bFGF. OBJECTIVE:To observe phenotypic changes and the bFGF protein expression after bFGF recombinant adenovirus was used to transfect rabbit bone marrow mesenchymal stem cells (BMSCs). METHODS:Passage 2 BMSCs were divided into three groups:Ad.bFGF-eGFP group, Ad.eGFP group and control group. Under a phase contrast microscope we observed the changes in cellmorphology. The expression of bFGF protein in BMSCs was determined by enzyme-linked immunosorbent assay (ELISA). The proliferative curve was detected by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT). RESULTS AND CONCLUSION:The transfected cells showed a uniform phenotype of fibroblasts. MTT colorimetric assay revealed that more proliferative activity of transfected BMSCs was shown in the Ad.bFGF-eGFP group than in the Ad.eGFP group and control group. ELISA results showed that expression of bFGF protein was higher in the Ad.bFGF-eGFP group than in the Ad.eGFP group and control group (P<0.05). BFGF recombinant adenovirus can induce the differentiation of BMSCs into fibroblasts, increase proliferative ability and promote the expression of bFGF protein.
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BACKGROUND:Cytotoxic T lymphocyte-associated antigen 4 is a newly discovered costimulatory molecule. It has been studied more in tumor and autoimmune diseases, less in the field of kidney transplantation. OBJECTIVE:To explore the role of cytotoxic T lymphocyte-associated antigen 4 in acute rejection after renal transplantation. METHODS:Fifty patients undergoing renal transplantation were divided into acute rejection group (20 cases) and stable graft function group (30 cases). Another 30 healthy persons served as control group. Blood samples were extracted from the peripheral blood. Cytotoxic T lymphocyte-associated antigen 4 was detected by enzyme linked immunosorbent assay and flow cytometry. RESULTS AND CONCLUSION:The expression of cytotoxic T lymphocyte-associated antigen 4 in the serum showed significant differences in the acute rejection group, stable graft function group and healthy control group (F=70.008 1, P=0.000 0), but showed no difference in peripheral blood lymphocytes of three groups (F=1.865 6, P=0.161 7). Compared with the healthy control group, the expression levels of cytotoxic T lymphocyte-associated antigen 4 in peripheral blood lymphocytes of acute rejection group and stable graft function group were significantly decreased (P=0.000 0). In addition, the acute rejection group had a lower cytotoxic T lymphocyte-associated antigen 4 expression than the stable graft function group (P=0.000 0). In renal transplant rejection, the expression of cytotoxic T lymphocyte-associated antigen 4 in serum was reduced, showing some correlation with acute rejection after renal transplnatation. Cytotoxic T lymphocyte-associated antigen 4 might be involved in the rejection.
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BACKGROUND:Pulmonary infection is the main complication after kidney transplantation, and its onset and morbidity may be related to conventional oral drugs after kidney transplantation. OBJECTIVE:To analyze the effect of methylprednisolone instead of prednisone on severe pulmonary infection after kidney transplantation. METHODS:Clinical data of 58 patients with severe pulmonary infection after kidney transplantation were retrospectively analyzed. First, according to the characteristics of post-onset patients and lung CT findings, broad-spectrum antibiotics and anti-fungal treatment were adopted, and subsequently targeted therapy, that is, withdrawal or adjustment of dosage and combination regimen of immunosuppressive agents, was employed depending on etiology, fungi and virus detection results. Among the 58 patients, 28 patients were injected methylprednisolone, and 30 patients took oral prednisone. Hyoxemia correction, support therapy and immune replacement therapy were applied. RESULTS AND CONCLUSION:Thirty-nine of 58 patients (67.2%) were positive for pathogens, including 7 cases of simple bacterial pneumonia, 4 cases of fungal pneumonia, 3 cases of simple cytomegalovirus infection, and 25 cases of mixed infections (5 cases of multiple bacterial infections, 17 cases of fungal and bacterial co-infections, and 3 cases of fungi, bacteria and cytomegalovirus co-infections). Patients subjected to methylprednisolone treatment spent shorter time to recover their temperature than those undergoing oral prednisone (P<0.05). In addition, creatinine fluctuation range in the methylprednisolone group was less than that in the prednisone group (P<0.05). The results showed that intravenous injection of methylprednisolone may accelerate absorption of inflammatory exudate in the lung and shorten treatment time.
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The article reviewed the research progress of ligustilide in recent years and elaborated its pharmacological functions and mechanisms in detail,especially in ischemic brain injury.Its mechanism includes reducing cerebral infarct volumes and improving neurobehavioral deficits,anti-oxidant and anti-apoptosis,antithrombotic activity,calcium channel blockers function,and effect on erythropoietin.Other pharmacological effects of ligustilide including inhibiting vascular smooth muscle cell proliferation,anti-inflammatory and analgesic effects,effects on LPS-induced endotoxic shock,inhibiting constriction effect,suppression of the central nervous system,and ameliorating the memory impairment induced by scopolamine and so on,are also introduced.Ligustilide has potential pharmacological value,which provides a reference for its further research and development.
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ObjectiveTo review the major complications in patients after transurethal electrovaporization of the prostate (TUVP) and transurethal plasmakinetic resection of the prostate (PKRP) retrospectively and to analyze the causes and management.MethodsClinical data of 92 cases of patients after TUVP and 226 cases after PKRP were reviewed retrospectively.The patients' relevant circumstances including subjective symptoms,objective indexes and the major long-term complications were followed up about 1-,3-,and 5-year after operation.Different therapeutic methods were chosen according to different causes of the complications.ResultsThere were no significant differences (P ＞ 0.05 ) between TUVP group and PKRP group in IPSS (7.3±2.8,7.2±2.5),QOL (2.6±0.7,2.7 ±0.5),Qmax[ (25.2±3.5),(25.5 ±3.8) ml/s] and PVR [(18.7 ±5.4),(17.8 ±6.3)ml].The incidences of bladder neck restriction was about 1.1％,3.3％,and 2.3％ after 1,3,and5 years in patients after TUVP,and 0.9％,2.7％,and 1.8％ after PKRP accordingly.For urethral stricture,it was about 3.3％,2.2％,and 1.1％ after TUVP,and 3.1％,2.2％,and 0.9％ after PKRP.For residual prostatic hyperplasia,it was about 1.1％,2.2％,and 4.5％ after TUVP,and 1.3％,2.7％,and 3.7％ after PKRP accordingly.ConclusionsTUVP and PKRP are effective and safe treatment options for BPH.The major long-term complications after TUVP and PKRP are bladder neck restriction,urethral stricture and residual prostatic hyperplasia.Regular and long-term follow-up is required for patients after TUVP and PKRP.Different therapeutic methods should be chosen according to different causes of the complications after operation.
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Objective To evaluate the efficacy and safety of solifenacin in the treatment of overactive bladder (OAB) syndrome in patients who have undergone transurethral resection of the prostate (TURP). Methods According to the Overactive Bladder Symptom Score (OABSS), 64 cases with OAB symptoms after TURP were randomly assigned into study and control groups with 32 cases in each group. Patients in the study group were treated with solifenacin (5 mg once daily) for a two week period beginning the first day after catheter removal. Patients in the control group were not treated with solifenacin. The mean urgency episodes, micturition episodes, nocturia, urge incontinence, volume voided per micturition, Qmax and OABSS scores were recorded on the 7th and the 14th day after catheter removal. Treatment-emergent adverse events with solifenacin in the study group were recorded and evaluated as well. All cases were followed-up for 8 weeks after catheter removal. Results There were statistically significant differences (P<0.01) in favor of the study group over the control group in the aspect of urgency, micturition episodes, nocturia, urge incontinence, volume voided per micturition and OABSS scores. The incidences of treatment related adverse events were 12.5% (4/32) in the study group with no serious adverse event observed. Conclusions Solifenacin is effective in the treatment of OAB syndrome after TURP and is well tolerated as well. Application of solifenacin should be recommended earlier after TURP.
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im To study the structure-function relationship of hTNFα . Methods We compared the cytotoxicity, receptor binding ability and toxicity in animal body of wild type(wt)hTNFα with its mutants including R2K-, N30S-, R32W-, L157F-hTNFα , and two multi-site mutants(R32W-L157F-hTNFα and R2K-N30S-R32W-L157F-hTNFα ). Results We found that the two multi-site mutants remained similar cytotoxicity to several human tumor cell lines as wild type hTNFα . However, their cytotoxicity to L929 cells were decreased sharply as compared with those of wt hTNFα . The two multi-site hTNFα mutants had lower binding activity with hTR75 than hTR55. We also found that compared with the wild type, the LD50 of the mutant R32W-L157F-hTNFα was decreased about 300 fold and the dose of mutant R2K-N30S-R32W-L157F-hTNFα resulted in 30％ death was 700 folds lower than LD50 of wt hTNFα . To certify the systematic toxicity of the mutant R2K-N30S-R32W-L157F-hTNFα , we assayed its toxicity to monkeys and found that its systematic toxicity was lower than that of wt hTNFα. Conclusion A 4-site mutants(R2k-N30S-R32W-L157F-hTNFα )of hTNFα is obtained, which the mutant may possess potential application value in clinical therapy.
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Purpose The aim is to establish fermentation process of new type recombinant human necrosis factor (nrhTNF)α.Methods The optimized host cell,culture medium and induction time were deternined according to the growth and expression specificity of nrhTNF on test tube and flask shaker. Then fed-batch culture was carried out on 5 L automatic fermentor.Results The expression of nrhTNF in fermentor could keep in a higher level of about 50% of total bacterial proteins when fermented to around A600nm 30，with a fermentation time of 12 h to 13 h.Conclusion A short cycle,high expression and stabilized fermentation process was established.
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Objective:To summarize the 10 year clinical experience of treating tibiafibular fractures with rectangle shaped intramedullary nails(RIN). Methods:From January 1987 to December 1996, 4 682 cases (3 278 male and 1 404 female) of tibiafibular fractures from 9 hospitals were treated with RIN . Three kinds of reduction methods including open reduction, semi open reduction and closed reduction were used during operation. Results:Results showed 2 173 cases (62.89%) got excellent result, 947 got good (27.40%), 214 got moderate (6.19%), 121 got poor (3.50%). The total healing rate was 90.29%. Conclusion:RIN has excellent biological characteristics which can provide a flexible interfixation when treating tibiafibular fractures, and the operation is simple, it also can be used for severe open fractures. RIN is one of the good techniques in treating tibiafibular fractures.
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Purpose To explore the possibility and the effect of therapeutic bronchial asthma by antisense oligonucleotid. Methods Based on the IL-5 cDNA sequence of mouse,a segment of antisense oligonucleotid was designed and synthetized.5′-labeling of antisense oligonucleotid was signed by T4 PNK in order that the efficiency of stearylamine liposome in transfe-ting antisense oligonucleotid can be evaluated. Astham model was duplicated with ovalbumin (OVA) absorbed to aluminum hydroxide. T lymphocytes of mice were separated by nylon fiber method,then T lymphocytes transfected a different content of antisense oligonucleotid with stearylamine phys. positive liposome were cultured respectively in order to observe the effect of antisense oligonucleotid on IL-5 produced by T lymphocytes. IL-5 levels in the supernatants of T lymphocytes culture were determined by ELISA. Results Stearylamine liposome could markedly increase the efficiency of antisense oligonucleotid transfection. The efficiency of antisense oligonucleotid transfection was the best at 1∶15 m/m(antisense oligonucleotid and SA liposome) and it was increased approximately 12 times.In healthy and asthma Balb/c mice, IL-5 was not detected in the supernatants of T lymphocytes culture without challenge with OVA.However,IL-5 was increased markedly in the supernatants of T lymphocytes culture challenged with OVA. After transfecting a different concentration antisense oligonucleotid, IL-5 levels in the supernatants of T lymphocytes culture were significantly lower than those in control cells without antisense oligonucleotid transfection. IL-5 levels decreased from (44.60±6.23) to (30.70±7.362),(17.20±6.181) and(8.16±2.34)pg/ml respectively. And IL-5 synthesis was inhibited by 31.17% , 61.43% and 81.7% respectively. Conclutions IL-5 synthesis could be obviously inhibited by antisense oligonucleotid and showed a markedly relation between quantitative and effect. It is supported that the production of IL-5 be inhibited through preventing the transcription of IL-5 from T lymphocytes. The study provides foundation for antisense gene therapeutic asthma.
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Objective To study the effects of ganoderma spore oil on Friend murine leukemia virus(Fr.MuLV)and its mechanism.Methods The BALB/C female mouse was infected with Fr.MuLV,body weight,spleen index,thymus index,the percentages of CD4+,CD8+,and CD4+/CD8+ were mea-sured.Results Compared to the control group,body weight extenuation,splenomegaly,and atrophy of thymus gland severely in the model group,the percentages of CD4+,CD8+,and CD4+/CD8+ were significantly decreased in the model.Body weight,spleen index,and thymus index(P
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Objective To establish HPLC fingerprint for Fructus Aurantii Immaturus.Methods The HPLC method was used with Diamonsil-C18 column (250 mm?4.6 mm,5 ?m),and a mixture liquid of acetonitrile-0.01% NaH2PO4 as mobile phase in a gradient elution.The HPLC fingerprint for 36 batches of Fructus Aurantii Immaturus was studied on their similarity,cluster,and principal components analyses.The common HPLC fingerprint of Fructus Aurantii Immaturus was established,which was studied with principal components analyses.Results Under the selected spectrum condition,the 36 batches of Fructus Aurantii Immaturus were classified into two groups based on the result of similarity,cluster,and principal components analyses.Conclusion This method is reasonable and reliable to the quality control of Fructus Aurantii Immaturus.
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Objective To compare the influence of different extraction technology on the pharmacodynamic actions of drug pair of Rhizoma Atractylodis Macrocephalae and Radix Paeoniae Alba.Methods The influences of water extraction,alcohol extraction,and SFE-CO2 extraction on pharmacodynamic actions of drug pair of Rhizoma Atractylodis Macrocephalae and Radix Paeoniae Alba were evaluated by mice intestine propulsive motility test,rats repelling acute gastric mucosa damage test,mice twisting test and hot-plate test.Results The paired drugs extracted by SFE-CO2 combining with water extraction or alcohol extraction had better pharmacological actions.Conclusion This study provides a scientific basis for optimizing the extraction technology of drug pair of Rhizoma Atractylodis Macrocephalae and Radix Paeoniae Alba.