4.Gray relational analysis for the effect of nano-drug features on drug absorption.
Jie YU ; Mao-sheng ZHENG ; Chao-ni XIAO ; Xiao-hui ZHENG
Acta Pharmaceutica Sinica 2015;50(9):1096-1100
Gray relational analysis is employed to analyze the effects of nano-drug features on phagocytosis of macrophages, including zeta potential, shape factor and scale size of drug particle, the correlation among such factors and macrophage cell is calculated. The results showed that the internalization processe is affected significantly by the zeta potential of the drug in the process of phagocytosis of macrophages, followed by the impact particle shape, and the effect of particle scale behaves relatively small.
Drug Carriers
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chemistry
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Humans
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Macrophages
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cytology
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Nanoparticles
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chemistry
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Particle Size
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Phagocytosis
5.The inhibitory effect of ozanimod on the growth and biofilm formation of Staphylococcus aureus
LIU Xiao-ming ; ZHANG Chao-qin ; XIAO Xiao-yin ; YU Zhi-jian ; ZHENG Jin-xin
China Tropical Medicine 2022;22(09):797-
Abstract: Objective To screening new compounds that can inhibit the growth and biofilm formation of Staphylococcus aureus. Methods Compounds that can inhibit the growth of Staphylococcus aureus were screened from the FDA approved drug library by 96 well plates. The absorbance value of 600 nm wavelength (OD600) was measured by Microplate Reader to detect the growth of Staphylococcus aureus planktonic cells in the culture supernatant. The minimum inhibitory concentration (MIC) of ozanimod against Staphylococcus aureus clinical isolates were detected by micro broth dilution method. The inhibitory effect of sub-inhibitory concentrations of ozanimod on the biofilm formation of Staphylococcus aureus was detected by crystal violet staining. Results This study found that ozanimod could significantly inhibit the growth of Staphylococcus aureus SA113 (screening reference strain), and the MIC was 25.00 μmol/L. The MIC of ozanimod against 119 clinical isolates of Staphylococcus aureus [65 isolates of methicillin sensitive (MSSA) and 54 isolates of methicillin resistant (MRSA)] was 12.50 or 25.00 μmol/L. The MIC50 and MIC90 of ozanimod against the 119 Staphylococcus aureus isolates all were 25.00 μmol/L. This study found that 6.25, 12.50, 25.00 μmol/L of ozanimod could significantly inhibit the biofilm formation of 2 MSSA and 2 MRSA. The sub-MIC concentration of ozanimod (12.50 μmol/L) could significantly inhibit the biofilm formation of 14 MSSA and 11 MRSA, but had no inhibitory effect on the growth of planktonic cells of these Staphylococcus aureus isolates. Conclusion Ozanimod can inhibit the growth of Staphylococcus aureus, including MRSA, and has good antibacterial activity. The sub-MIC concentration of ozanimod could significantly inhibit the biofilm formation of Staphylococcus aureus.
6.Large-scale expansion of clinical-grade human adipose-derived stem cells using the extracellular matrix
Yuehan SU ; Chao WEI ; Pinlei LV ; Yun CAO ; Yun QIU ; Qing ZHENG ; Shudong XIAO ; Zheng WANG
Chinese Journal of Tissue Engineering Research 2014;(10):1521-1531
BACKGROUND:Large-scale expansion of undifferentiated and multipotential adipose-derived stem cells using serum-free culture system is a difficult issue to be resolved. OBJECTIVE:To establish an in vitro culture system combined with the extracellular matrix in order to investigate the efficiency, effectiveness and security of extracellular matrix on expanding adipose-derived stem cells. METHODS:In vitro isolated adipose-derived stem cells were seeded in traditional two-dimensional plastic plates and extracellular matrix-coated plates supplemented with serum-free medium respectively. After in vitro expansion, total cellnumber, expression of cellsurface markers, cellsenescence degree and multipotent differentiation ability (adipogenic, osteoblastic and chondrogenic differentiation) of adipose-derived stem cells cultured under both conditions were detected and compared. Moreover, the clinical safety of adipose-derived stem cells expanded in extracellular matrix-coated plates was investigated. RESULTS AND CONCLUSION:Total cellnumber of passage 5 adipose-derived stem cells cultured in extracellular matrix-coated plates was 10 times more than that in traditional two-dimensional plastic plates. Flow-cytometric analysis showed that adipose-derived stem cells cultured with extracellular matrix expressed stem cellsurface markers. cellular senescence examination showed that almost al of passage 15 adipose-derived stem cells cultured with extracellular matrix showed no aging, while most passage 5 adipose-derived stem cells cultured by the two-dimensional system aged and lost their proliferation ability. Multidirectional induction of adipose-derived stem cells showed that passage 15 adipose-derived stem cells cultured with extracellular matrix could stil differentiate into adipocytes, osteoblasts and chondrocytes as passage 5 adipose-derived stem cells did, which performed much better than the induced differentiations of passage 5 adipose-derived stem cells cultured by the two-dimensional system. Karyotype analysis and in vivo invasion experiment insured the clinical safety of adipose-derived stem cells expanded with extracellular matrix. Al above results suggest a safe and more efficient expansion system of extracellular matrix for clinical application using the serum-free culture system combined with extracellular matrix.
7.FTIR fingerprint spectrograms of traditional Chinese medicine Marsdenia tenacissima.
Chao LI ; Sheng-Chao YANG ; Qiao-Sheng GUO ; Kai-Yan ZHENG ; Ping-Li WANG ; Xiang-Zeng XU ; Xue-Feng XIAO
China Journal of Chinese Materia Medica 2014;39(17):3311-3315
In this paper, Fourier transform infrared spectroscopy fingerprint analysis of Marsdenia tenacissima samples was used to develop a reliable method of tracing the geographical origins. Forty-eight samples from four provinces of China were analyzed by FTIR. We analyzed and characterized the fingerprints in both the full spectrum peaks and characteristic peaks, then the principal component analysis and the cluster analysis were carried out. The results of fingerprint analysis, correlation analysis, principal component analysis and cluster analysis can identify the geographic origins correctly, which verified and supplemented each other; the identification results and the actual location showed a high degree of consistency, namely the lower the space distance, the greater the similarity of different samples. These results revealed the obvious superiority and practical value in comparison to the more tedious and time-consuming wet chemistry method normally used. Using appropriate metrology methods can trace the geographical source correctly. The M. tenacissima materials from the region of Maguan should be considered as genuine medicinal materials taking into account the good quality.
China
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Cluster Analysis
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Drugs, Chinese Herbal
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analysis
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classification
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standards
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Geography
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Marsdenia
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chemistry
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classification
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Medicine, Chinese Traditional
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Principal Component Analysis
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Quality Control
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Reproducibility of Results
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Spectroscopy, Fourier Transform Infrared
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methods
8.Diagnostic value of digital breast tomosynthesis plus full-field digital mammography in breast disease
Chao YOU ; Yajia GU ; Weijun PENG ; Tingting JIANG ; Xiaojing ZHENG ; Qin XIAO
Chinese Journal of Radiology 2015;(2):94-98
Objective To investigate the diagnostic value of digital breast tomosynthesis (DBT) plus full-field digital mammography (FFDM) in benign and malignant breast disease.Methods The prospective study enrolled 46 consenting women with a clinical suspicious of breast lesion confirmed by pathology. All the patients underwent DBT, FFDM and dedicated breast magnetic resonance image (DB-MRI) examinations before surgery or biopsy. To observe the manifestation of breast disease in different examinations according to breast imaging reporting and data system (BI-RADS), and to evaluate the visibility grade of disease between DBT plus FFDM and FFDM alone. Using receiving operating characteristic(ROC) and Z test to analysis the diagnostic efficiency of three examinations.Results Forty-six cases included 14 benign cases and 32 malignant cases. Compared with FFDM, 35 cases showed increased visibility in DBT plus FFDM, that the malignant lesion manifested mass with speculated and architectural distortion(23/27) and the benign lesion manifested mass with halo sign(6/8). Eleven cases didn't show increase visibility in both DBT plus FFDM and FFDM alone. The ration of BI-RADS 0 in FFDM was 23.9%(11/46), meanwhile in other two examination were 0. The ration of BI-RADS 3 in DBT plus FFDM was the lowest(6.5%, 3/46). When the lesion in DBT plus FFDM showed mass or speculated, in MRI showed similar morphology with mass or mass-like enhancement. According pathology as the gold standard, excluded 11 cases with BI-RADS 0 in FFDM, there was significant statistical difference between DBT plus FFDM and FFDM alone in 35 cases (Z=2.132, P=0.033) to differentiate benign and malignant breast disease, the area of ROC curve were respectively 0.955, 0.878. There is no statistical difference between DBT plus FFDM and MRI in 46 cases (Z=0.502, P=0.616), the area of ROC curve were respectively 0.887, 0.908.Conclusion DBT combined with FFDM can improve both detection of breast lesion and diagnostic efficiency, while the performance value between DBT and MRI are equal.
9.Effects of rosiglitazone on antagonizing insulin resistance and adipose liver damnification in hyperlipemic rats
Linying ZHENG ; Jingqiang PAN ; Jieru LIN ; Liuying XIAO ; Boping LI ; Chao HAN
Chinese Journal of Tissue Engineering Research 2007;11(16):3189-3192,3200
BACKGROUND: Some experiments indicated that applying rosiglitazone on diabetic animals lacking of insulin could not increase insulin and lower blood glucose obviously, which showed that rosiglitazone did not stimulate the excretion of rosiglitazone. The action of rosiglitazone in improving insulin resistance and the effects on the functions of liver and kidneys need more investigations.OBJECTIVE: To investigate whether rosiglitazone can improve the insulin resistance of rats with hyperlipemia, and analyze the possible mechanism.SETTINGS: Guangzhou Hospital of Traditional Chinese Medicine; Guangzhou Institute of Traditional Chinese Medicine and Materia MedicaDESIGN: A stratified randomized controlled animal trial.MATERIALS: Sixty-four Sprague-Dawley (SD) rats (Batch No. 2002A024), SPF grade, half male and half female,weighing 150 to 180 g, aged 6 to 8 weeks were purchased from Guangdong Medical Experimental Animal Center.Normal feed (total quantity of heat 6.9 kJ/g) was enriched with 23% protein, 53% carbohydrate and 5% fat. High fat emulsion (total quantity of heat 15.5 kJ/g) was enriched with 200 g/L lard, 200 g/L cholesterol, 10 g/L bile salt ox,200 g/L propylene glycol, 200 g/L tween-80. High fat and sugar feed (total quantity of heat 21.0 kJ/g) was enriched with 15% protein, 51% carbohydrate and 30% fat after adding 100 g/L glucose, 200 g/L lard and 100 g/L yolk powder then mixing and baking. Rosiglitazone was from GlaxoSmithKline Co Ltd. (Tianjin) (5 mg/tab, Batch No.02110012). Gliclazide was from Servier International and Tianjin Hua Jin Pharmaceutical Factory (100 mg/tab, Batch No.00232).METHODS: The experiment was carried out in Guangzhou University of Traditional Chinese Medicine from April to July in 2003. ① Sixty-four Sprague-Dawley rats, 16 of which were randomly sampled as the normal control group and had been fed with normal feed for 6 weeks. The others were modeled after medical literatures, each one was administered with high fat emulsion (10 mL/kg) by gavage once a day for 14 days. Rats whose FBG≥6.1 mmol/L or 2hBG≥7.8 mmol/L were selected, randomized into 3 groups according to body mass and blood glucose, i.e., negative control (model)group, rosiglitazone group and gliclazide group, there were 16 rats in each group. Except the normal control group, rats in the rosiglitazone group and gliclazide group were gavaged with rosiglitazone for 5 mg/kg and gliclazide for 100 mg/kg respectively, and those in the model group were gavaged with distilled water. All of the rats were fed with high-fat feed once a day for 28 days. From the 21st day, high fat emulsion was added once a day for 7 days. After fasting for 18 hours from the last administration, all the rats were recorded for FBG and administered dextrose 2.78 mol/10 mL .kg or dextrose and drug mixture 10 mL/kg by body mass. Two hours'later, 2hBG was recorded. ② Blood samples were collected from orbital plexus and serum was prepared for detecting the biochemical indexes and immunological indexes in serum, i.e., fasting serum glucose(FSG), total cholesterol (TC), triglyceride (TG), high density lipoprotein cholesterol (HDL-C), low density lipoprotein cholesterol (LDL-C), alanine aminotransferase (ALT), aspartate aminotransferase (AST),blood urea nitrogen (BUN), creatinine (Cr), tumor necrosis factor alpha (TNF-α) and fasting insulin (FINS). The insulin sensitivity index (ISI) was calculated: ISI=ln [1/ (FINS content×FBG content)]. After the rats were killed, their liver suspension was prepared for measuring the levels of TG, superoxide dismutase (SOD), glutathione (GSH) and malondialdehyde (MDA).MAIN OUTCOME MEASURES: ① FBG and 2hBG; ② FSG, blood lipids, TNF-α, FINS and ISI in serum; ③ TG, GSH, SOD and MDA in liver cells; ④ ALT, AST, BUN and Cr in serum. RESULTS: ① Results of FBG and 2hBG: The FBG and 2hBG in the rosiglitazone group [(3.2±0.3), (6.3±1.2) mmol/L]were lower than those in the modelcontrol group [(3.8±0.5), (8.1±2.1) mmol/L, P < 0.01]. The FBG in the gliclazide group [(3.3±0.7) mmol/L] was lower than that in the model control group. ② Results of FSG, blood lipids, TNF-α, FINS and ISI: The FSG, TNF-α and FINS in the rosiglitazone group were (4.2±1.2) mmol/L, (246±45) μg/L and (133±45) pmol/L respectively, which were lower than those in the model control group [(6.6±1.5) mmol/L, (294±65) μg/L, (264±76) pmol/L,P < 0.05-0.01], whereas ISI was higher than that in the model control group (-6.33±0.46, -7.46±0.95, P < 0.01). The FSG and TNF-α in the gliclazide group [(4.1±1.1) mmol/L, (251±62) μg/L] were lower than those in the model control group (P < 0.05-0.01). ③ Results of TG content, GSH deposit, SOD activity and MDA content in liver cells: The TG and MDA contents in liver cells in the rosiglitazone group [(1.00±0.38), (40±17) mmol/g] were lower than those in the model control group [(2.40±0.60), (171±63) mmol/g, P< 0.01], the GSH deposit and SOD activity [(51±14) mg/g, (583.45±50.01 ) nkat/g] were higher than those in the model control group [(2.40±0.60) mg/g, (450.09±66. 68) nkat/g, P < 0.05-0.01].The TG and MDA contents in the gliclazide group [(1.20±0.38), (100±30) mmol/g] were lower than those in the model control group, whereas the GSH deposit [(46±15) mg/g] was higher than that in the model control group. ④ Results of ALT, AST, BUN and Cr in serum: The serum contents of BUN and Cr in the rosiglitazone group [(14.3±3.8) mmol/L,(33±9) μmol/L] were lower than those in the model control group [(19.2±5.6) mmol/L, (45±13) μmol/L, P < 0.05].CONCLUSION: Both rosiglitazone and gliclazide can improve the insulin resistance induced by high fat feed.Rosiglitazone is superior to gliclazide in decreasing the high insulin level, decreaseing serum levels of BUN and Cr,improving reduced GSH deposit and enhancing SOD activity.
10.Immunosuppresive, antioxidative, anti-inflammatory and analgesic effect of Xiahuoluo pills
Jingqiang PAN ; Liuying XIAO ; Dan ZHANG ; Chao HAN ; Jieru LIN ; Linying ZHENG ; Boping LI
Chinese Journal of Tissue Engineering Research 2006;10(47):183-188
BACKGROUND: Xiaohuoluo pill can expel pathogenic wind, remove dampness and activate collaterals. It is used for treatment of Bi-syndrome due to wind-cold-dampness, pain and numbness in limbs.OBJECTIVE: To observe the pharmacological effect of Xiaohuoluo pills on secondary immune response, specific immunity (including cellular immunity and humoral immunity), non-specific immunity [including complement 3(C3), mononuclear phagocyte system (MPS) and red blood cell (RBC)adhesion function] and free radical injury as well as pain and many other inflammations in mice.DESIGN: A randomized controlled stratified trial.SETTING: Guangzhou Institute of Traditional Chinese Medicine and Chinese Materia Medica; Department of Pharmacy, Guangzhou Hospital of Traditional Chinese Medicine.MATERIALS: Totally 628 NIH and ICR mice of 6 to 8 weeks were involved in this trial. Xiaohuoluo pills (components: Dannanxing, Zhichuanwu, Zhicaowu, Dilong, Ruxiang and so on; Chenli Pharmaceutical Factory,Guangzhou; Brach No. 19980612) were used in this trial. Rabbit antimouse immunoglobulin G (IgG) and C3 antiserum reagent kit (Guangzhou Institute of Medicine and Health) and reagent kit for measuring the antioxidizing activity of superoxide dismutase (SOD) and the level of malondialdehyde (MDA) (Jiancheng Institute of Bioengineering, Nanjing) were used.METHODS: This trial was carried out in the Guangzhou Institute of Traditional Chinese Medicine and Chinese Materia Medica; Department of Pharmacy, Guangzhou Institute of Medicine and Health during September 1998 to December 1999. ① To observe the suppressive effect of Xiaohuoluo pills on cock red blood cell (CRBC)-induced secondary immune response: Eight-four ICR mice, male and female in half, were selected.Twenty of 84 mice served as blank controls; The other 64 mice were intraperitoneally injected with cyclophos-phamide (CY) of 0.2 g/kgonce. On the 4th and 12th days, CRBC was intraperitoneally injected into the mice twice to induce immunoenhancing pathological models to form secondary immune response. Mice served as blank controls were intraperitoneally injected with the same volume of normal saline; The immunoenhanced mice were assigned into 3 groups by a lot: CY group (n=20, CY, 40 mg/kg, intragastric administration, I.g.), Xiaohuoluo pills (n=21, Xiaohuoluo pills suspension, 5.54 g/kg, I.g.) and model group (n=20, distilled water, the same volume as other groups, I.g.); once a day within 7 successive days. 19 days later, the levels of serum IgG and C3 were measured with single immunodiffusion method, and the level of circulating immune compound (CIC) was measured with polyethylene glycol precipitation method. ② To observe the suppressive effect of Xiaohuoluo pills on delayed type hypersensitivity (DTH): Fifty-four ICR mice, male and female in half, were selected. On the 1st day, the mice were sensitized by subcutaneous injection of 10 g/L 2,4-dinitrofluorobenzen e (DNFB) of 50 μL for each. On the 4th day, the sensitized mice were assigned into 3 groups by a lot: Prednisone group (n=18, prednisone, 0.01 g/kg, I.g.), Xiaohuoluo pills (n=18, Xiaohuoluo pills suspension, 5.54 g/kg, I.g.), model group (n=18, distilled water, the same volume as other groups, I.g.), all once a day within 7 successive days. 11 days later, 10 g/L DNFB of 25μL was spread on the right ear of each mouse in each group. The swelling degree was calculated 24 hours later (The mass difference between right ear and left ear). ③ To observe the suppressive effect of Xiaohuoluo pills on immune adhesion function of RBC of mouse: Thirty-six NIH mice, male and female in half, were selected and assigned into 3 groups by a lot: CY group (n=12, CY, 20 mg/kg,I.g.), Xiaohuoluo pills (n=12, Xiaohuoluo pills suspension, 5.54 g/kg, I.g.)and blank control group (n=12, distilled water, the same volume as other groups, I.g.), once a day within 7 successive days. 7 days later, blood was taken from the orbit of mice for calculating the rosette rate of RBC-C3b receptor and the rosette rate of RBC immune compound. ④ To observe the suppressive effect o20 Mg/kg, I.g.),Xiaohuoluo pills group (Xiaohuoluo pills suspension, 5.54 g/kg, I.g.) , once a day within 7 successive days; IgM-type hemolytic concentration (HC50)was measured at 2 hours after the last administration on the 7th day [ (Sample absorption / Absorption at HC50 of CRBC) ×diluted time]. The levels of serum C3 and MDA and the activity of SOD were measured according to the method from corresponding reagent kit. ⑥ To observe the suppressive effect of Xiaohuoluo pills on agar granulation tissue hyperplasia in mice:Fifty-nine NIH mice were selected and given subcutaneous injection of 20 g/L agar of 0.5 mL for each. 24 hours later, the mice were assigned into 3 groups by a lot: diclofenac group (diclofenac, 10 mg/kg,I.g..), Xiaohuoluo pills group (Xiaohuoluo pills suspension, 5.54 g/kg, I.g.) and model group (distilled water, the same volume as other groups, I.g.), once a day within 7 successive days; On the 8th day, the mice were sacrificed. The hyperplasiainhibiting effect was presented in the form of the mass of agar granulation tissue in one kilogram body mass ⑦ To observe the suppressive effect of Xiaohuoluo pills on acetic distortion reaction: Sixty-three NIH mice were se lected and assigned into 3 groups by a lot: diclofenac group (diclofenac,50 mg/kg, I.g.), Xiaohuoluo pills group (Xiaohuoluo pills suspension, 5.54 g/kg,I.g.) and model group (distilled water, the same volume of other groups, I.g.),once a day within 2 successive days. At 2 hours after the last administration, the mice were given intraperitoneal injection of 0.1 mol/L acetic acid of 0.2 mL for each one. The times of distortion of mice within 20 minutes were counted. ⑧ To observe the effect of Xiaohuoluo pills on the acute exudative inflammation evoked by dimethylbenzene, croton oil and carrageenan, and the level of prostaglandin E in the inflammatory exudates:Totally 219 NIH mice were selected and assigned into 3 groups by a lot:diclofenac group (diclofenac, 50 mg/kg, I.g.), Xiaohuoluo pills group (Xiaohuoluo pills suspension, 5.54 g/kg, I.g.) and model group (distilled water, the same volume of other groups, I.g.) once a day within 2 successive days. At 2 hours after the last administration, dimethylbenzene of 25 μL was spread on the right ear for 20 minutes, or croton oil of 25 μL was also spread on the right ear, 4 hours later, the swelling of right ear was calculated (mass of right ear-mass of left ear). 10 g/L carrageenan of 20 μL was subcutaneously injected into the right foot, 3 hours later, the swelling degree was calculated (The difference of right foot and left foot); and the level of prostaglandin E in the inflammatory exudates was measured. ⑨ t test(t' test for heteroscedasticity) was used for comparing the difference in measurement data among groups.MAIN OUTCOME MEASURES: Pharmacological effect of Xiaohuoluo pills on secondary immune response, specific immunity, non-specific immunity and free radical injury as well as pain and many other inflammations in mice.RESULTS: Totally 628 NIH and ICR mice were involved in result analysis. ① The level of IgG and CIC of mice in the model group was significantly higher than that in the other 3 groups respectively (P < 0.01),while the level of C3 was significantly lower than that in the other 3 groups (P < 0.05 to 0.01). ② The swelling degree of mice in the diclofenac group and Xiaohuoluo pills group was significantly lower than that in the blank control group respectively ( both P < 0.01). ③ The rosette rate of RBC-C3b receptor and RBC immune compound in the blank control group was significantly higher than that in the other 2 gro ups respectively (P < 0.01). ④ The phagocytic index (K value )in the diclofenac group and Xiaohuoluo pills group was significantly lower than that in the blank control group, respectively (both P < 0.01).⑤ IgM-type HC50 and the level of serum MDA of CY group and Xiaohuoluo pills group were obviously lower than those in the immune control group (P < 0.01),while the level of C3 was higher than that of immune control group, there was no significant difference in the activity of serum SOD between CY group or Xiaohuoluo pills group and immune control group (P > 0.05). ⑥The ratio of agar granulation tissue mass to body mass in the diclofenac group or Xiaohuoluo pills group was significantly lower than that in the model group(P < 0.01).⑦ The times of distortion of mice within 20 minutes in the diclofenac group or Xiaohuoluo pills group were signifi cantly less than those of model group(P < 0.01,0.05).⑧The ear swelling degree of dimethylbenzene-induced inflammatory models and croton oil-induced inflammatory models,and foot swelling degree of carrageenan-induced acute inflammatory models as well as the level of prostaglandin E in the inflammatory exudates in the diclofenac group were significantly milder or lower than those in the model group(P < 0.05 to 0.01),and the level of prostaglandin E in the inflammatory exudates in the Xiaohuoluo pills group was significantly lower than that in the model group (P < 0.01).CONCLUSION: Xiaohuoluo pills possess pharmacological effects of immunosuppression, anti-proliferative inflammation, analgesia and antioxidation.