Effect of Haimiding on the functioning of red cell membrane of FC and H22 tumor-bearing mice

Table 1 Red cell membrane preparation procedure.

Whole blood
↓	Centrifuged at 3000 r/min for 10 min, rinsed twice with 1:3 saline, and suspended in 1:1 saline
Red cell suspension
↓	5p 8.4 phosphoric buffer solution (1:30) was added at 4 °C and hemolysis was allowed to proceed for 1 h.
Hemolyzed solution of red cells
↓	Centrifuged at 2000 r/min at 4 °C for 40 min; the supernatant was discarded; the residue was rinsed 3 times and then suspended in 5p8.4 phosphoric acid buffer solution.
Membrane suspension

Table 2 Steps for measuring the activity of the sialic acid content on the surface of the red cell membrane.

Reagent	Blank	1	2	3	4	5	6	7
N-acetyl neuraminic acid Standard solution (mL)	0	0.05	0.1	0.2	0.3	0.4	0.5	0.8
N-acetyl neuraminic acid (mg)	0	5	10	20	30	40	50	80
Distilled water (mL)	1	0.95	0.9	0.8	0.7	0.6	0.5	0.2
Bialsche reagent (mL)	1	1	1	1	1	1	1	1
Boil for 12 min in water bath, then cool for 3 min with ice water
Pentanol (mL)	5	5	5	5	5	5	5	5
Shaken thoroughly and then centrifuged for 10 min at 1 000 r/min; took the pentanol phase for colorimetric analysis at a wavelength 569 nm

Table 3 Steps for measuring the activity of the oxidoreductase NADH-cell pigment C.

Reagent (mL)	No saponin added (-S)		Saponin added (+S)
	Blank	Sample	Blank	Sample
0.6 mg protein/mL Membrane suspension	0.6	0.6	0.6	0.6
PBS (5p8.4)	2.4	2.2	2.2	2
0.1% saponin	/	/	0.2	0.2
5 mmol/L K3Fe(CN)6	/	0.1	/	0.1
Let the solution stand for 10 min, added NADH, and recorded the result after 1 min 6 mmol/L NADH	/	0.1	/	0.1

Table 4 Effect of Haimiding on the immunological adhesiveness of red cells of FC mice to tumor cells (mean±SD).

Group	Number of animals (n)	Route of administration	Dosage (mg/kg)	Rate of wreaths (%)
Control	10	Ip		36.36±5.42
Saline	10	Ip	Equal volume	10.46±3.55
5-Fu	10	Ip	25	8.41±2.56
Haimiding	10	Ip	50	28.96±4.41b

^bP<0.01 vs the saline group.

Table 5 Effect of Haimiding on the immunological adhesiveness of red cells of H₂₂ mice to tumor cells mean±SD.

Group	Dosage (mg/kg)	Route of administration	No. of animals (n)	Rate of wreaths (%)
Control		Ip	10	36.36±5.42
Saline	Equal volume	Ip	10	5.10±2.28
5-Fu	25	Ip	10	5.96±3.15
Haimiding	50	Ip	10	25.68±4.13b

^bP<0.01 vs the saline group.

Table 6 Effect of Haimiding on the fluidity of intact red cell membrane of FC mice.

Group	Number ofanimals (n)	Route ofadministration	Dosage (mg/kg)	Fluorescencepolarization (P)	Micro viscosity, h(P)	Membrane lipidfluidity (LFU)
Control	10		Equal volume	0.2006±0.0728	1.7557±0.9616	7.4472±2.3273
Saline	10	Ip	25	0.2438±0.0122	1.9458±0.2367	4.3134±1.4378
5-Fu	10	Ip	100	0.2169±0.0269	1.8183±0.3799	6.0234±1.9430a
Haimiding	10	Ip	50	0.215±0.0242b	1.796±0.1414a	6.1913±1.9348a
Haimiding	10	Ip	25	0.210±0.0137b	1.701±0.2063a	6.4966±1.9687b
Haimiding	10	Ip		0.2203±0.0338	1.7330±0.4404	5.7670±1.6962a

^aP<0.05, ^bP<0.01 vs the saline group.

Table 7 Effect of Haimiding on the fluidity of intact red cell membrane of H₂₂ mice.

Group	Number ofanimals (n)	Route ofadministration	Dosage (mg/kg)	Fluorescencepolarization (P)	Micro viscosity, h(P)	Membrane lipidfluidity (LFU)
Control	10			0.2006±0.0728	1.7557±0.9616	7.4478±2.3273
Saline	10	Ip	Equal volume	0.2430±0.03017	2.1378±0.6186	4.3559±1.4051
5Fu	10	Ip	25	0.2275±0.0182	1.9760±0.6738	5.2606±1.5472
Haimiding	10	Ip	100	0.2331±0.0211	2.0819±0.1772	4.9153±1.5360
Haimiding	10	Ip	50	1.2775±0.4164b	11.0370±3.4489b	0.1770±0.0567b
Haimiding	10	Ip	25	0.2111±0.0223a	1.8176±0.2106	6.4776±2.0895a

^aP<0.05, ^bP<0.01 vs the saline group.

Table 8 Effect of Haimiding on the membrane protein content in the red cells of FC mice.

Group	Number of animals (n)	Route of administration	Dosage (mg/kg)	Membrane protein content %
Control	10	Ip	Equal volume	0.281±0.051
Saline	10	Ip	25	0.332±0.094
5-Fu	10	Ip	100	0.278±0.041
Haimiding	10	Ip	50	0.241±0.037a
Haimiding	10	Ip	25	0.187±0.042b
Haimiding	10	Ip		0.296±0.040

^aP<0.05, ^bP<0.01 vs the saline group.

Table 9 Effect of Haimiding on the membrane protein content in the red cells of H₂₂ mice.

Group	Number of animals (n)	Route of administration	Dosage (mg/kg)	Membrane protein content %
Control	10		Equal volume	0.281±0.051
Saline	10	Ip	25	0.298±0.088
5-Fu	10	Ip	100	0.219±0.089
Haimiding	10	Ip	50	0.237±0.050
Haimiding	10	Ip	25	0.143±0.064b
Haimiding	10	Ip		0.1890.094a

^aP<0.05, ^bP<0.01 vs the saline group.

Table 10 Effect of Haimiding on the sialic acid content on the surface of the red cell membrane of FC mice.

Group	Number of animals (n)	Route of administration	Dosage (mg/kg)	A
Control	10	Ip		0.183±0.068
Saline	10	Ip	Equal volume	0.087±0.026
5-Fu	10	Ip	25	0.065±0.030
Haimiding	10	Ip	100	0.139±0.049b
Haimiding	10	Ip	50	0.148±0.032b
Haimiding	10	Ip	25	0.095±0.028

^bP<0.01 vs the saline group.

Table 11 Effect of Haimiding on the sialic acid content on the surface of the red cell membrane of H₂₂ mice.

Group	Number of animals (n)	Route of administration	Dosage (mg/kg)	A
Control	10	Ip		0.183±0.068
Saline	10	Ip	Equal volume	0.062±0.027
5-Fu	10	Ip	25	0.064±0.034
Haimiding	10	Ip	100	0.119±0.040b
Haimiding	10	Ip	50	0.125±0.044b
Haimiding	10	Ip	25	0.081±0.023

^bP<0.01 vs the saline group.

Table 12 Effect of Haimiding on the sealability of the red cell membrane of FC mice.

Group	Number of animals (n)	Route of administration	Dosage (mg/kg)	Sealability (%)
Control	10	Ip		70.6±4.05
Saline	10	Ip	Equal volume	54.6±2.21
5-Fu	10	Ip	25	52.0±3.39
Haimiding	10	Ip	100	63.5±3.11b
Haimiding	10	Ip	50	65.9±2.60b
Haimiding	10	Ip	25	58.4±2.05b

^bP<0.01 vs the saline group.

Table 13 Effect of Haimiding on the sealability of the red cell membrane of H₂₂ mice.

Group	Number of animals (n)	Route of administration	Dosage (mg/kg)	Sealability (%)
Control	10	Ip		70.2±2.23
Saline	10	Ip	Equal volume	53.7±2.30
5-Fu	10	Ip	25	54.2±3.27
Haimiding	10	Ip	100	62.2±2.71b
Haimiding	10	Ip	50	61.4±3.44b
Haimiding	10	Ip	25	57.6±2.54b

^bP<0.01 vs the saline group.