Application of multiple strategies to enhance oleanolic acid biosynthesis by engineered Saccharomyces cerevisiae.

Oleanolic acid and its derivatives are widely used in the pharmaceutical, agricultural, cosmetic and food industries. Previous studies have shown that oleanolic acid production levels in engineered cell factories are low, which is why oleanolic acid is still widely extracted from traditional medicinal plants. To construct a highly efficient oleanolic acid production strain, rate-limiting steps were regulated by inducible promoters and the expression of key genes in the oleanolic acid synthetic pathway was enhanced. Subsequently, precursor pool expansion, pathway refactoring and diploid construction were considered to harmonize cell growth and oleanolic acid production. The multi-strategy combination promoted oleanolic acid production of up to 4.07 g/L in a 100 L bioreactor, which was the highest level reported.

A role for the cerebellum in motor-triggered alleviation of anxiety.

Physical exercise is known to reduce anxiety, but the underlying brain mechanisms remain unclear. Here, we explore a hypothalamo-cerebello-amygdalar circuit that may mediate motor-dependent alleviation of anxiety. This three-neuron loop, in which the cerebellar dentate nucleus takes center stage, bridges the motor system with the emotional system. Subjecting animals to a constant rotarod engages glutamatergic cerebellar dentate neurons that drive PKCδ+ amygdalar neurons to elicit an anxiolytic effect. Moreover, challenging animals on an accelerated rather than a constant rotarod engages hypothalamic neurons that provide a superimposed anxiolytic effect via an orexinergic projection to the dentate neurons that activate the amygdala. Our findings reveal a cerebello-limbic pathway that may contribute to motor-triggered alleviation of anxiety and that may be optimally exploited during challenging physical exercise.

The Meridian Tropism and Classification of Red Yeast Rice Investigated by Monitoring Dermal Electrical Potential.

Red yeast rice is a traditional Chinese medicine and food that has been purported to color food, ferment, and lower cholesterol. In order to study the antioxidative capacity of red yeast rice and the effects on electrical potential difference (EPD) of 12 acupuncture meridians, the pH value, oxidation reduction potential (ORP), ABTS, FRAP, T-SOD, and particle size distribution of red yeast rice were analyzed. 20 volunteers were recruited and randomly divided into two groups, the red yeast rice group (10 g red yeast rice and 40 g water) and control CK group (50 g water). The left 12 acupuncture meridians' EPD was real-time monitored. Samples were taken at the 10th minutes. The whole procedure continued for 70 minutes. It is shown that the pH value of the red yeast rice was 4.22, the ORP was 359.63 mV, the ABTS was 0.48 mmol Trolox, the FRAP was 0.08 mmol FeSO4, the T-SOD was 4.71 U, and the average particle size was 108 nm (7.1%) and 398.1 nm (92.9%). The results of 12 acupuncture meridians' EPD showed that the red yeast rice can significantly affect the EPD of stomach, heart, small intestine, and liver meridians.

Effect of Electroacupuncture at Zusanli (ST36) on Sepsis Induced by Cecal Ligation Puncture and Its Relevance to Spleen.

BACKGROUND: Acupuncture at Zusanli (ST36), Quchi (LI11), and Tianshu (ST25) is commonly used in septic patients by traditional Chinese physicians. The protective effect of acupuncture at ST36 on the intestinal barrier is associated with Cholinergic Anti-Inflammatory Pathway (CAIP). However, its detailed mechanism and whether acupuncture at LI11 and ST25 have similar effects to ST36 remain unclear. AIM: To explore the effects of electroacupuncture (EA) at ST36, LI11, and ST25 on septic rats and investigate the role of the spleen in the treatment of EA at ST36. METHODS: A septic rat model caused by cecal ligation and puncture (CLP) and a postsplenectomy (SPX) CLP rat model were established. Rats were divided into nine groups depending on different treatments. Serum levels of TNF-α, IL-10, D-lactic acidosis (D-LA), double amine oxidase (DAO), and T-lymphocyte subgroup level in intestinal lymph nodes were compared. RESULTS: EA could not improve the 2-day survival of CLP rats. For CLP rats, EA at ST36 and LI11 significantly decreased the levels of TNF-α, IL-10, DAO, and D-LA in serum and normalized intestinal T-cell immunity. For SPX CLP rats, EA at ST36 failed to reduce serum concentrations of TNF-α, IL-10, and D-LA but increased the values of CD3+CD4+/CD3+CD8+ cells and Treg/Th17 cells. CONCLUSIONS: EA at ST36 and LI11, respectively, could alleviate inflammation reaction, protect the intestinal barrier, and maintain intestinal T-cell function in septic rats. Spleen participated in the protective effect of EA at ST36 in sepsis.

Effects of TAT-SOD at Acupoints on Essential Hypertension by Monitoring Meridians Electrical Potential.

OBJECTIVES: To investigate the effect on essential hypertension of the topical application of TAT-Cu, Zn-superoxide dismutase (TAT-SOD) at left acupoint Zusanli (ST 36), and to observe whether the change of electrical potential difference (EPD) can be related to the change of blood pressure. METHODS: Sixteen patients with essential hypertension and 16 healthy subjects were included in the study. EPD between the left acupoints of Yanglingquan (GB 34) and Qiuxu (GB 40) was firstly screened out for the EPD detection. An intracellular superoxide quenching enzyme, TAT-SOD, was topically applied to the acupoint ST 36 within an area of 1 cm2 once a day, and the influence on EPD was investigated. The dosage applied to TAT-SOD group (n=8) was 0.2 mL of 3000 U/mL TAT-SOD cream prepared by adding purified TAT-SOD to a vehicle cream, while placebo group (n=8) used the vehicle cream instead. The left acupoints of Yanglingquan (GB 34) and Qiuxu (GB 40) were selected for EPD measurement after comparing EPD readings between 5 acupoints on each of all 12 meridians. RESULTS: EPDs between the left acupoints of GB 34 and GB 40 for 16 patients of essential hypertension and 16 healthy subjects were 44.9±6.4 and 5.6±0.9 mV, respectively. Daily application of TAT-SOD for 15 days at ST 36 of essential hypertension patients significantly decreased systolic blood pressure (SBP) and diastolic blood pressure (DBP) of 179.6 and 81.5 mm Hg to 153.1 and 74.1 mm Hg, respectively. Responding to the change in blood pressure, EPD between the left acupoints of GB 34 and GB 40 also declined from 44.4 to 22.8 mV with the same trend. No change was observed with SBP, DBP and EPD between the left acupoints of GB 34 and GB 40 with the daily application of the placebo cream. CONCLUSION: Enzymatic scavenging of the intracellular superoxide at ST 36 proved to be effective in decreasing SBP and DBP. The results reconfirm the involvement of superoxide anions and its transportation along the meridians, and demonstrate that EPD between acupoints may be an indicator to reflect its functioning status. Moreover, preliminary results suggest a close correlation between EPD and blood pressure readings, implying a possibility of using EPD as a sensitive parameter for blood pressure and to monitor the effect of antihypertensive treatment.

Salvianolic acid B promotes neural differentiation of induced pluripotent stem cells via PI3K/AKT/GSK3ß/ß-catenin pathway.

Salvianolic acid B (Sal B), a water-soluble component mainly extracted from the traditional Chinese medicine Salvia miltiorrhiza, has potential anti-inflammatory, anti-oxidative and anti-apoptotic actions to protect neural cells. Here, we explore the effects and mechanisms of Sal B on the promotion of differentiation of induced pluripotent stem cells (iPSCs) into neural stem cells (NSCs), then further into neurons. During the processes of neural differentiation of iPSCs, Sal B or a phosphatidylinositide 3 kinase (PI3K) inhibitor (LY294002) were added to the medium. Sal B substantially improved proliferation of iPSC-derived NSCs and neurons. Furthermore, Sal B significantly stimulated PI3K/AKT/GSK3 ß/ß-catenin pathway. However, LY294002 attenuated the Sal B-induced increase. Therefore, these outcomes suggest that Sal B markedly enhances neural differentiation of iPSCs via the PI3K/AKT/GSK3ß/ß-catenin pathway.

[Effect of Medicated-catgut Embedding at "Changqiang" (GV 1) on Mechanical Pain Threshold and P 38 MAPK Expression in Spinal Cord Tissue in Anus Incisional Pain Rats].

OBJECTIVE: To observe the effect of medicated-catgut embedding at "Changqiang"(GV 1) on regional pain reaction and expression of p 38 MAPK in the dorsal horn of spinal cord in anus incisional pain rats, so as to explore its analgesic mechanism. METHODS: Forty male SD rats were randomly divided into control, model, GV 1-embedding and sham acupoint embedding groups (n=10 rats in each group). The anus incisional pain model was established by making a radial incision (about 10 mm length) at the left lithotomy position of the anus with a surgical knife, and the mechanical pain threshold (PT) was measured by using a Von Frey before and 4, 8, 12, 24 h after operation. The medicated-catgut (about 12.5 mm length/kg body weight) was implanted in the subcutaneous tissue of GV 1 region. The immunoactivity of p 38 MAPK was determined by immunohistochemistry. RESULTS: Compared with the control group, the mechanical PTs were significantly decreased 4, 8, 12 and 24 h after operation both at the site of incision and about 15 mm proximal to the site of incision in the model group (P<0.05), and the immunoactivity of phosphorylated (p)-p 38 MAPK in the superficial layer of dorsal horns of lumbar spinal cord was significantly increased(24 h)after operation(P<0.05). Compared with the model group, the PTs were significantly increased 8, 12 and 24 h after operation at the site of incision, and 12 h and 24 h at the site about 15 mm proximal to the incision region (P<0.05), and the immunoactivity level of p-p 38 MAPK was significantly down-regulated in the GV 1-embedding group (P<0.05). No significant changes were found in the PT and p-p 38 MAPK immunoactivity levels in the sham acupoint embedding group (P<0.05). CONCLUSIONS: Medicated-catgut embedding at "Changqiang"(GV 1) has an analgesic effect in anus incisional pain model rats, which may be related to its effect in down-regulating the expression of p 38 MAPK in the dorsal horn of lumbar spinal cord.

Topical Application of TAT-Superoxide Dismutase in Acupoints LI 20 on Allergic Rhinitis.

Reactive oxygen species are products of cellular metabolism and assigned important roles in biomedical science as deleterious factors in pathologies. In fact, some studies have shown that the therapeutic benefits of taking antioxidants were limited and the potential for therapeutic intervention remains unclear. New evidences showed that ROS have some ability of intercellular transportation. For treating allergic rhinitis, as a novel intracellular superoxide quencher, TAT-SOD applied to acupoints LI 20 instead of directly to nasal cavity can be used to test that. TTA group apply TAT-SOD cream prepared by adding purified TAT-SOD to the vehicle cream to acupoints LI 20, while placebo group used the vehicle cream instead. TTN group applied the same TAT-SOD cream directly to nasal cavity three times daily. Symptom scores were recorded at baseline and days 8 and 15. For the overall efficacy rate, TTA group was 81.0%, while placebo group was 5.9% and TTN was 0%. Malondialdehyde levels decreased observably in TTA group, and superoxide dismutase, catalase, and glutathione peroxidase levels remained basically unaffected. Enzymatic scavenging of the intracellular superoxide at acupoints LI 20 proved to be effective in treating allergic rhinitis, while no improvement was observed with the placebo group and TTN group.

A Systematic Review of Rhubarb (a Traditional Chinese Medicine) Used for the Treatment of Experimental Sepsis.

Sepsis is a global major health problem in great need for more effective therapy. For thousands of years, Rhubarb had been used for various diseases including severe infection. Pharmacological studies and trials reported that Rhubarb may be effective in treating sepsis, but the efficacy and the quality of evidence remain unclear since there is no systematic review on Rhubarb for sepsis. The present study is the first systematic review of Rhubarb used for the treatment of experimental sepsis in both English and Chinese literatures by identifying 27 studies from 7 databases. It showed that Rhubarb might be effective in reducing injuries in gastrointestinal tract, lung, and liver induced by sepsis, and its potential mechanisms might include reducing oxidative stress and inflammation, ameliorating microcirculatory disturbance, and maintaining immune balance. Yet the positive findings should be interpreted with caution due to poor methodological quality. In a word, Rhubarb might be a promising candidate that is worth further clinical and experimental trials for sepsis therapy.

Monitoring the Effects of Acupoint Antioxidant Intervention by Measuring Electrical Potential Difference along the Meridian.

Previous studies suggest that superoxide anions are possibly traveling along acupuncture meridians. The electrical potential difference (EPD) between acupoints may be related to the movement. To test the above hypothesis, we conducted a study investigating the effects of acupoint antioxidant interventions on the meridian EPD. Firstly, ST39 (L) and ST44 (L) were screened out for the EPD detection along the stomach meridian, and ST36 (L) was selected for interventions including acumassage with the control cream, as well as the TAT-SOD cream for 30 minutes, or injection with reduced glutathione sodium. The EPD between ST39 and ST44 was recorded for 80 minutes and measured again 48 h later. While the EPD increased during the acumassage, the acumassage with TAT-SOD cream and the glutathione injection generated waves of EPD increased, indicating the migration or removal from the visceral organ of a greater quantity of superoxide. Remarkably lower EPD readings 48 h later with both antioxidant acupoint interventions than the mere acumassage imply a more complete superoxide flushing out due to the restored superoxide pathway at the acupoint after interventions. The results confirm superoxide transportation along the meridians and demonstrate a possibility of acupoint EPD measurement as a tool to monitor changes in the meridians and acupoints.

Protective Effects and Mechanisms of Salvianolic Acid B Against H2O2-Induced Injury in Induced Pluripotent Stem Cell-Derived Neural Stem Cells.

Induced pluripotent stem cells (iPSCs) have the potential to differentiate into neural lineages. Salvianolic acid B (Sal B) is a commonly used, traditional Chinese medicine for enhancing neuroprotective effects, and has antioxidant, anti-inflammatory, and antiapoptotic properties. Here, we explore the potential mechanism of Sal B in protecting iPSC-derived neural stem cells (NSCs) against H2O2-induced injury. iPSCs were induced into NSCs, iPSC-derived NSCs were treated with 50 µM Sal B for 24.5 h and 500 µM H2O2 for 24 h. The resulting effects were examined by flow cytometry analysis, quantitative reverse-transcription polymerase chain reaction, and western blotting. Upon H2O2 exposure, Sal B significantly promoted cell viability and stabilization of the mitochondrial membrane potential. Sal B also visibly decreased the cell apoptotic ratio. In addition, Sal B markedly reduced expression of matrix metalloproteinase (MMP)-2 and -9, and phosphospecific signal transducer and activator of transcription 3 (p-STAT3), and increased the level of tissue inhibitor of metalloproteinase (TIMP)-2 in iPSC-derived NSCs induced by H2O2. These results suggest that Sal B protects iPSC-derived NSCs against H2O2-induced oxidative stress. The mechanisms of this stress tolerance may be attributed to modulation of the MMP/TIMP system and inhibition of the STAT3 signaling pathway.

Inspissated bile syndrome in an infant with citrin deficiency and congenital anomalies of the biliary tract and esophagus: identification and pathogenicity analysis of a novel SLC25A13 mutation with incomplete penetrance.

Biallelic mutations of the SLC25A13 gene result in citrin deficiency (CD) in humans. Neonatal intrahepatic cholestasis caused by citrin deficiency (NICCD) is the major CD phenotype in pediatrics; however, knowledge on its genotypic and phenotypic characteristics remains limited. The present study aimed to explore novel molecular and clinical characteristics of CD. An infant suspected to have NICCD as well as her parents were enrolled as the research subjects. SLC25A13 mutations were investigated using various methods, including cDNA cloning and sequencing. The pathogenicity of a novel mutation was analyzed bioinformatically and functionally with a yeast model. Both the infant and her father were heterozygous for c.2T>C and c.790G>A, while the mother was only a c.2T>C carrier. The novel c.790G>A mutation proved bioinformatically and functionally pathogenic. The infant had esophageal atresia and an accessory hepatic duct, along with bile plug formation confirmed by laparoscopic surgery. However, the father seemed to be healthy thus far. The findings of the present study enrich the genotypic and phenotypic characteristics of CD patients, and provided clinical and molecular evidence suggesting the possible non-penetrance of SLC25A13 mutations and the likely involvement of this gene in primitive foregut development during early embryonic life.

Effects of Salvia miltiorrhiza on neural differentiation of induced pluripotent stem cells.

ETHNOPHARMACOLOGICAL RELEVANCE: Salvia miltiorrhiza, a well-known traditional Chinese medicine, is commonly used to treat some neurological diseases because of its anti-oxidative, anti-inflammatory and anti-apoptotic properties. We investigate whether Salvia miltiorrhiza can improve the differentiation of induced pluripotent stem cells (iPSCs) into neurons in vitro, and promote iPSCs-derived neural stem cells survival, integrate, and differentiation after their transplantation to the ischemic brain tissues. MATERIALS AND METHODS: Induced pluripotent stem cells were used to differentiate into neural stem cells, and further into neurons in induction medium with various concentrations of Salvia miltiorrhiza. The effects were assessed by immunofluorescence staining, quantitative reverse transcription-polymerase chain reaction (qRT-PCR) and Western blotting. iPSC-derived neural stem cells were transplanted into the brains of rats with middle cerebral artery occlusion, immunofluorescence staining was used to evaluate survival, integrate, and differentiation of grafted cells, the functional recovery of the animals was tested by the Longa scores and spontaneous motor activity. RESULTS: Salvia miltiorrhiza (5µg/ml) significantly increased the gene and protein expression of Nestin compared with that in other groups. Microtubule-associated protein 2 (MAP2) expression in induction medium with 5µg/ml Salvia miltiorrhiza was significantly higher than that in the control group. After cells transplantation into the ischemic brain, more grafted MAP2(+) cells were found in Salvia miltiorrhiza-treated rats than others at 7 days. Salvia miltiorrhiza-treated rats showed the most remarkable functional recovery at 7 and 14 days. CONCLUSION: Salvia miltiorrhiza induces differentiation of induced pluripotent stem cells to differentiate into neurons efficiently. The plant provides neuroprotection to implanted cells and improves functional recovery after their transplantation to the ischemic brain tissues.

[Factors influencing the spatial variability in soil respiration under different land use regimes].

In order to investigate the factors influencing the spatial variability in soil respiration under different land use regimes, field experiments were performed. Soil respiration and relevant environment, vegetation and soil factors were measured. The spatial variability in soil respiration and the relationship between soil respiration and these measured factors were investigated. Results indicated that land use regimes had significant effects on soil respiration. Soil respiration varied significantly (P < 0.001) among different land use regimes. Soil respiration rates ranged from 1.82 to 7.46 micromol x (m2 x s)(-1), with a difference of 5.62 micromol x (m2 x s)(-1) between the highest and lowest respiration rates. Soil organic carbon was a key factor controlling the spatial variability in soil respiration. In all, ecosystems studied, the relationship between soil respiration and soil organic carbon content can be described by a power function. Soil respiration increased with the increase of soil organic carbon. In forest ecosystem, the relationship between soil respiration and diameter at breast height (DBH) of trees can be explained by a natural logarithmic function. A model composed of soil organic carbon (C, %), available phosphorous (AP, g x kg(-1)) and diameter at breast height (DBH, cm) explained 92.8% spatial variability in soil respiration for forest ecosystems.

[Postoperative pain of hemorrhoid treated with thread embedding at Changqiang (GV 1)].

OBJECTIVE: To compare the therapeutic effect of postoperative pain of hemorrhoid treated with thread embedding at Changqiang (GV 1) and Indometacin suppository. METHODS: Eighty cases were randomly divided into an acupoint catgut-embedding group (group A) and an Indometacin group (group B), forty case in each group. In group A, thread embedding at Changqiang (GV 1) was applied, and Indometacin suppository was put into rectum in group B. The pain intensity after 6, 12, 24, 72 hours of treatment and the the maximum scores of VAS in 24 hours (T24max VAS) of 1-3 days of postopration were evaluated with pain Visual Analogue Scale (VAS); the quantity demanded of analgesic and the adverse reaction were observed. RESULTS: The scores of VAS after 6, 12, 24, 72 hours of treatment in group A were obviously lower than those in group B (all P < 0.05); the T24max VAS scores in group A were obviously lower than those in group B (all P < 0.05). Six cases in group A and 14 cases in group B were treated with non-steroidal anti-inflammatory drug as well. The total effective rate of 87.5% (35/40) in group A was superior to that of 62.5% (25/40) in group B, indicating the statistically significant differences between two groups (P < 0.05). There was no obvious adverse reaction in group A; in group B, headache, dizziness, nausea, vomiting, distention of chest and palpitation appeared, accounting for 11 cases. CONCLUSION: The therapeutic effect of postoperative pain of hemorrhoid treated with thread embedding at Changqiang (GV 1) is favorable, it can reduce the quantity of analgesic, the adverse reaction, its effect is better than that of Indometacin suppository.

[Mechanism study of anti-influenza effects of Radix Isatidis water extract by red blood cells capillary electrophoresis].

OBJECTIVE: To research the mechanism of the inhibition effects of BWE on cell attachment of influenza virus by capillary electrophoresis. METHOD: The morphologic difference of red cells after treating with BWE infected by influenza virus was detected with microscope, capillary electrophoresis and HA. RESULT: The pretreatment of the normal cells with BWE inhibited the attachment of influenza to the cells, while no meaningful inhibition was observed when influenza virus was pretreated before being inoculated to cells. CONCLUSION: The results indicate that the inhibition effects of BWE on cell attachment of influenza virus may be an important mechanism of anti-influenza activity of Radix Isatidis Extracts.

Mechanism study of anti-influenza effects of Radix Isatidis water extract by red blood cells capillary electrophoresis / 中国中药杂志

<p><b>OBJECTIVE</b>To research the mechanism of the inhibition effects of BWE on cell attachment of influenza virus by capillary electrophoresis.</p><p><b>METHOD</b>The morphologic difference of red cells after treating with BWE infected by influenza virus was detected with microscope, capillary electrophoresis and HA.</p><p><b>RESULT</b>The pretreatment of the normal cells with BWE inhibited the attachment of influenza to the cells, while no meaningful inhibition was observed when influenza virus was pretreated before being inoculated to cells.</p><p><b>CONCLUSION</b>The results indicate that the inhibition effects of BWE on cell attachment of influenza virus may be an important mechanism of anti-influenza activity of Radix Isatidis Extracts.</p>

[The modulation of pilose antler extract (PAE) on the proliferation of rat osteogenic cells UMR-106].

OBJECTIVE: To investigate the modulation of pilose antler extract (PAE) on rat osteogenic cells UMR-106 in vitro. METHOD: Component P2 of PAE was isolated by Sephacryl S-200HR gel filtration chromatography. The proliferative effects of P2 and other components isolated by Sephacryl S-200HR on UMR-106 cells were investigated by MTT assay. RESULT: The P2 could significantly increase the proliferation rate of osteogenic cells. When the protein concentration of P2 was between 0.972 mg x L(-1) and 97.2 mg x L(-1), it could inhibit the proliferation of UMR-106 cells. But while the concentration was equal to or greater than 97.2 mg x L(-1), the P2 could increase the proliferation rate of cells, especially 477.92% at 9.72 g x L(-1), which was approximated to 499.62% of PAE. The molecular weight of the P2 was about 59 kDa determined by SDS-PAGE. On the other hand, inhibition was also observed in the sample of the P3, P4 and P5. CONCLUSION: Those regulative factors in PAE which have different molecular weight can affect the proliferation of UMR-106 cells two-wayly. And this adjustment also relies on the dose of those factors. This finding may help us to understand the possible mechanism of Chinese traditional medicine from animal materials.

The modulation of pilose antler extract (PAE) on the proliferation of rat osteogenic cells UMR-106 / 中国中药杂志

<p><b>OBJECTIVE</b>To investigate the modulation of pilose antler extract (PAE) on rat osteogenic cells UMR-106 in vitro.</p><p><b>METHOD</b>Component P2 of PAE was isolated by Sephacryl S-200HR gel filtration chromatography. The proliferative effects of P2 and other components isolated by Sephacryl S-200HR on UMR-106 cells were investigated by MTT assay.</p><p><b>RESULT</b>The P2 could significantly increase the proliferation rate of osteogenic cells. When the protein concentration of P2 was between 0.972 mg x L(-1) and 97.2 mg x L(-1), it could inhibit the proliferation of UMR-106 cells. But while the concentration was equal to or greater than 97.2 mg x L(-1), the P2 could increase the proliferation rate of cells, especially 477.92% at 9.72 g x L(-1), which was approximated to 499.62% of PAE. The molecular weight of the P2 was about 59 kDa determined by SDS-PAGE. On the other hand, inhibition was also observed in the sample of the P3, P4 and P5.</p><p><b>CONCLUSION</b>Those regulative factors in PAE which have different molecular weight can affect the proliferation of UMR-106 cells two-wayly. And this adjustment also relies on the dose of those factors. This finding may help us to understand the possible mechanism of Chinese traditional medicine from animal materials.</p>