Biological Evaluation for Quality Control of Water Extract of Qingjin Huatantang Based on Phagocytic and Secretory Functions of Macrophages / 中国实验方剂学杂志

Objective:To establish a method for evaluating the biological activity of water extract lyophilized powder of Qingjin Huatantang based on the phagocytic and secretory functions of macrophages， and to control the quality of this formula from the biological activity level. Method:The phagocytic and inflammation models of RAW264.7 macrophages were established， the inhibition rates of water extract lyophilized powder of Qingjin Huatantang on interleukin-6 （IL-6） secretion and phagocytic index of neutral red of RAW264.7 macrophages were chosen as indicators to investigate the biological activity of Qingjin Huatantang， and the biological limit was searched. Result:The optimal inoculation density of RAW264.7 macrophages was 3×10⁵ pcs/mL， and the concentration of lipopolysaccharide （LPS） was 1 mg·L^-1 after treatment for 24 h. When the concentration was 500 mg·L^-1， water extract lyophilized powder of Qingjin Huatantang had no toxicity and no obvious promotion effect on the proliferation of RAW264.7 macrophages， and at this concentration， the phagocytosis of RAW264.7 macrophages for neutral red was significantly promoted， the phagocytic index was >113%. In addition， the lyophilized powder had a significant and stable inhibitory effect on IL-6 secretion of RAW264.7 macrophages induced by LPS， the inhibitory rate was >45%. Conclusion:Combined with the anti-inflammatory and immunomodulatory effects of Qingjin Huatantang， this study establishes an <italic>in vitro </italic>biological limit method for evaluating the quality of water extract of Qingjin Huatantang based on the phagocytic and secretory functions of RAW264.7 macrophages， and 500 mg·L^-1 was confirmed as the limit concentration. Under the limit concentration， Qingjin Huatantang water extract can significantly promote the phagocytic index of macrophages or significantly inhibit the secretion of IL-6 of RAW264.7 macrophages induced by LPS， which can be judged as qualified.

Evaluation Of Immunomodulatory Activity Of Diosgenin In Rats

Background: Theimmune system is intrinsic to health. Modulation of the immune responses to alleviate the diseases by using herbal plants has been of interest for many years. Diosgenin, a naturally occurring steroid saponin mainly present in the seeds of fenugreek (Trigonella foenum graecum)and in the root tubers of wild yams (Dioscorea vil-losa). Activation of specific and nonspecific immunity results in stimulation of immune response. Diosgenin has the positive effects on both specific and nonspecific immunity.Aim: To study the immunomodulatory activity of Diosgen-in in rats. Method: The suspension of Diosgenin wasgiven orally at the dosage level of 50, 100 and 150 mg/kg for 21 days in a rat. The immunomodulatory activity on specific and non-specific immunity was studied by haemagglutina-tion antibody (HA) titer, delayed type hypersensitivity (DTH) response and carbon clearance test. Immunosuppres-sion in a rat was induced by using Cyclophosphamide (100 mg/kg, p.o.). Sheep red blood cells (SRBCs) were used as antigen (0.1ml 20% SRBCs) in haemagglutinating antibody titer and delayed type hypersensitivity response methods. Result: Diosgenin exhibited significant increase in the production of antibody titer in response to SRBC antigen. A significant increase in both primary and secondary HA titer was observed in immunosuppressed group treated with Diosgenin when compared with negative control. A significant increase in the DTH response was observed in immu-nosuppressed animals treated with Diosgenin, pre-sensitized with SRBCs antigen. Diosgenin exhibited significant in-crease in phagocytic index against control group, indicating the stimulation of the reticuloendothelial system. Con-clusion: The study indicates that Diosgenin triggers stimulatory effect on specific and nonspecific immune response. The immunostimulant effect of Diosgenin could be attributed due to its saponin glycoside.

Enhancement effect of Superfine Cordyceps militarisPowder on immune functions of mice / 吉林大学学报(医学版)

Objective:To investigate the effect of Superfine Cordyceps militarisv Powder on the immune functions of the mice,and to provide basis for improving the utilizable ratio of Cordyceps militaris.Methods:A total of 40 mice were randomly divided into negative control group,low dose (0.166 5 g·kg-1),middle dose (0.333 3 g·kg-1),and high dose (0.999 9 g·kg-1) of Superfine Cordyceps militaris Powder groups,and there were 10 mice in each group.The mice in different doses of Superfine Cordyceps militaris Powder groups were administered with the Superfine Cordyceps militaris Powder for 30 d by intragastric infusion respectively,whereas,the mice in negative control group were administered with the same volume of water for 30 d by intragastric infusion.The body weights,the spleen indexes and thymus indexes of the mice in various groups were measured;the lymphocyte transformation abilities of the mice in various groups were observed by lymphocyte transformation experiment;the levels of delayed-type hypersensitivity reaction of the mice were examined with toe incrassation;the number of antibody forming cells,phagocytic rate and phagocytic index of chicken erythrocytes phagocytized by peritoneal macrophages of the mice were detected.Results:Compared with negative control group,the body weights,the spleen indexes a nd thymus indexes of the mice in different doses of Superfine Cordyceps militaris Powder groups had no significant differences (P>0.05).The lymphocyte transformation abilities of the mice in middle and high doses of Superfine Cordyceps militarisPowder groups were higher than that in negative control group(P<0.05).The levels of delayed-type hypersensitivity reaction of the mice in middle and high doses of Superfine Cordyceps militaris Powder groups were higher than those in low dose of Superfine Cordyceps militaris powder group and negative control group(P<0.05).The numbers of antibody forming cells of the mice in low,middle and high doses of Superfine Cordyceps militaris Powder groups were higher than that in negative control group(P<0.05).The phagocytic rates and phagocytic indexes of chicken erythrocytes phagocytized by peritoneal macrophage of the mice in low,middle and high doses of Superfine Cordyceps militaris Powder groups were higher than that in negative control group(P<0.05).Conclusion:Superfine Cordyceps militaris Powder can strengthen the immune functions of the mice,and the recommended doses are 0.333 3 and 0.999 9 g·kg-1.

Carissa congesta Wight and Benincasa hispida (Thunb.) Cogn. as budding immunomodulatory agents.

Carissa congesta and Benincasa hispida are well-known medicinally important plants associated with diabetes, inflammation, protozal infections and cancer. Here, we emphasized up on the immunomodulatory potential of these plants as the source of lupeol, β-sitosterol and ursolic acid. Petroleum ether extracts of C. congesta roots and B. hispida seeds were subjected to acute toxicity studies. They were screened for its immunomodulatory prospective in rats by Haemagglutination Antibody (HA) titre and Delayed-Type Hypersensitivity (DTH) response using Sheep Red Blood Cells (SRBCs of-0.5×109) as antigens. Carbon Clearance test (Phagocytic Index) was estimated by Indian ink suspension. Complete Freund’s Adjuvant (CFA) induced arthritis model interpretation was done by paw edema, kene joint erosion (transverse section), body weights, arthritic index and biochemical levels (RBC, WBC and Hb levels). Both the extracts were found to be therapeutically safe up to 5000 mg/kg. Dosage of 100 mg/kg was not satisfactory; and 500 and 250 mg/kg showed significant immunostimmulation (HA Titre) and immunosuppression (DTH response, 48 h). Benincasa hispida seed and Carissa congesta root extracts showed phagocytic Index of 0.0163±0.003, 0.0145±0.003 and 0.0183±0.003, 0.0176±0.003 at 250 mg/kg and 500 mg/kg, respectively. CFA model revealed that the B. hispida seed and C. congesta root extracts decreased paw volume, knee joint erosion, increased body weights and biochemical parameters with an arthritic index of 1.31±0.12, 1.44±0.15 and 1. 16±0.09, 1.36±0.13 at 250 mg/kg and 500 mg, respectively. The results were interpreted by One-way ANOVA followed by Dunnett test. Extracts showed relevance as promising immunostimulators as compared to control.

Immunomodulatory Effect of Water Extract of Stachytarpheta jamaicensis (L.) Vahl.

Stachytarpheta jamaicensis (L)Vahl traditionally is used to treat several diseases, such as urinary tract infections, rheumatic, and pharyngitis. The objective of this study was to test immunomodulatory effect of water extract of S. jamaicensis (L.) Vahl. herbs. Immunomodulatory assay of water extract of S. jamaicensis herbs have been examined in male Balb/c mice using carbon clearance assay. The doses of the extract were 62.5, 125, and 250 mg/kg bw. Parameter of immunomodulatory effect was phagocytic indexes after seven days administration of the extract. The result showed thatwater extract of S. jamaicensis at doses of 62.5 and 250 mg/kg bw had lower phagocytic index compared to Zymosan A, but higher phagocytic indexes compared to Methylprednisolone. The water extract of S. jamaicensismay be considered as an immunosuppresant agent.

Induction of phagocytic activity and nitric-oxide production in natural populations of Trypanosoma Cruzi I and II from the state of Paraná, Brazil / Indução da atividade fagocitária e produção de óxido nítrico numa população natural de Trypanosoma cruzi I e II do Estado do Paraná, Brasil

Twelve strains of Trypanosoma cruzi isolated from wild reservoirs, triatomines, and chronic chagasic patients in the state of Paraná, southern Brazil, and classified as T. cruzi I and II, were used to test the correlation between genetic and biological diversity. The Phagocytic Index (PI) and nitric-oxide (NO) production in vitro were used as biological parameters. The PI of the T. cruzi I and II strains did not differ significantly, nor did the PI of the T. cruzi strains isolated from humans, triatomines, or wild reservoirs. There was a statistical difference in the inhibition of NO production between T. cruzi I and II and between parasites isolated from humans and the strains isolated from triatomines and wild reservoirs, but there was no correlation between genetics and biology when the strains were analyzed independently of the lineages or hosts from which the strains were isolated. There were significant correlations for Randomly Amplified Polymorphic Deoxyribonucleic acid (RAPD) and biological parameters for T. cruzi I and II, and for humans or wild reservoirs when the lineages or hosts were considered individually.

Doze cepas de Trypanosoma cruzi isoladas de reservatórios silvestres, triatomíneos e de pacientes chagásicos crônicos do Estado do Paraná, Brasil, classificadas como Tc I e II foram usadas para avaliar a correlação entre genética e diversidade biológica. Índice fagocítico (IF) e produção de óxido nítrico (ON) in vitro foram os parâmetros biológicos utilizados. O IF de cepas T. cruzi I e II não diferiram significativamente assim como o IF de cepas isoladas de humanos, triatomíneos ou de reservatórios silvestres. Há diferença estatística na inibição da produção de ON entre T. cruzi I e II e entre parasitos isolados de humanos e de cepas isoladas de triatomíneos e reservatórios silvestres, mas não foi observada correlação entre genética e biologia quando as cepas foram analisadas independentemente da linhagem ou hospedeiros das quais elas foram isoladas. Observou-se correlação significativa para amplificação aleatória do DNA polimórfico e parâmetros biológicos de Tc I ou II e para os seres humanos ou reservatório silvestre quando linhagens ou hospedeiros são consideradas separadamente.

Immunomodulatory effect of Moringa oleifera Lam. extract on cyclophosphamide induced toxicity in mice.

Immunomodulatory effect of ethanolic extract (50%) of M. oleifera leaves (MOE) has been studied in normal and immunosuppressed mice models. Different doses of MOE i.e. 125, 250 and 500 mg/kg body weight of mice were administered orally for 15 days. Cyclophosphamide at a dose of 30 mg / kg body weight was administered orally for the next 3 days. On day 16 and 19, hematological parameters like white blood cell (WBC) count, red blood cell (RBC) count, haemoglobin level (Hb), percent neutrophils and organ weight were recorded. Effect of MOE on phagocytic activity of mice macrophages was determined by carbon clearance test. MOE showed significant dose dependent increase in WBC, percent neutrophils, weight of thymus and spleen along with phagocytic index in normal and immunosuppressed mice. The results indicate that MOE significantly reduced cyclophosphamide induced immunosuppression by stimulating both cellular and humoral immunity.

Effect of surgical stress on neutrophil function.

The present study was undertaken to find out the effect of surgical stress on nonspecific immune response. Twenty patients posted for various elective surgeries participated in the study (male : 17, female : 3, age : 43.4±2 yrs). The blood samples were taken preoperatively (4 to 6 days prior to surgery) and the following parameters were assessed: phagocytic index of neutrophils, avidity index of neutrophils and percentage of neutrophils in differential count. These were compared with the respective parameters assessed in the blood samples taken 24 hours after surgery. There was a significant (P=0.0001) decrease in the phagocytic index of neutrophil and a significant (P=0.003) increase in the percentage of neutrophils in differential count in the postoperative blood samples. However, the avidity index did not show a significant change. It could be tentatively concluded that surgical stress causes depression of nonspecific immunity in the early postoperative period.

Pharmacokinetic Studies of Radix Acanthopanacis Senticosis Suspension / 中药新药与临床药理

Objective To observe the pharmacokinetic feature of Radix Acanthopanacis Senticosis suspension(RASS). Methods The research was conducted with pharmacodynamic endpoint approach and pharmacodynamic approach and adopted with phagocytic index as pharmacological parameter. Results Pharmacodynamic endpoint approach: K_(el)=0.2d~(-1), K_a=2.5 d~(-1), AUC(S_0)=10.4g?kg~(-1)?d, VRT=19.3d~2.Pharmacodynamic approach: K_(el(ef))=0.1d~(-1), K_(a(ef))=2.9d~(-1), AUC(S_0)_((ef))=62.7(%)?d, VRT_((ef))=43.2d~2. The data obtained by pharmacodynamic approach differed from that by pharmacodynamic endpoint approach, indicating the delay of drug effect. Conclusion Pharmacokinetic Data of RASS are first obtained in this article, which show that pharmacodynamic endpoint approach and pharmacodynamic approach is feasible.