In the present paper, it is reported that immunelogic event during unset of experimehtaleallrgic encephalomyelitis (EAE) is studied by sevaeral immunological parameters, including thymectomy at birth in Guinea Pigs, identification of various cellular and humoral immunolo-gic functions. It is observed that thymectomy at birth suppressed onset of EAE and severityof EAE is related to cellular immunologic function but not to humoral immunity. All ofthese results indicated that cell-mediated immunity is responsible for the onset of EAE.

Toll-like receptors is a super-family of pathogen recognition-receptors discovered in recent years.During the process of ischemia-reperfusion, the Toll-like receptor 4 (TLR4) combines with lipopolysaccharide and many endogenous ligands such as high mobility group protein B1, heparan sulfate, and fibrinogen. Through the myeloid differentiation protein 88 -dependent and -independent signaling pathways, the products induce the release of inflammatory cytokine-mediated inflammatory response, leading to injuries. ω-3 polyunsaturated fatty acids, by inhibiting the signal pathway activation and target gene expression of TLR4, can influence the function of many immune cells and regulate the body's inflammatory response and immune function. This article reviews the function of TLR4 during ischemia-reperfusion injury and the possible interventional role of ω-3 polyunsaturated fatty acids.

In cooperation with a District Health Station of Beijing, 13 nurseries were selected for the study of the practice of diet planning. At the beginning, the health workers of the selected nurseries -were gathered together at the station to attend a short-term class. Lectures were given on the basic knowledge of child nutrition, the methods of dietary survey and diet planning, i.e. using the proper amount of the cheaper but more nutritive foods to make essentially balanced diets according to the RDA. The health workers of the nurseries meet and comunicate once a month at the health station to calculate the monthly food consumption and the average daily intake of the nutrients of the children in the past month and to make plan for the next month. The body weight of the children were measured every month and the height at three months intervals. The data collected in 12 months were summarized. The results indicated that the yearly average intake of most nutrients by the children increased and their collective growth level in weight and height increased correspondingly during the one year period of diet planning without any increase of the boarding expense of the children.

OBJECTIVE:To establish a HPLC method for the determination of main component and the related substances in pirenoxinum natricum eye drops. METHODS: A Inertsil C8-3 column was used with pH2.2 perchloric acid-methanol (35∶65) as mobile phase at a flow of 1.0 mL?min-1. The detection wavelength was 230 nm and the sample size was 10 ?L. RESULTS: Pirenoxinum natricum and the related substances could be baseline separated. The calibration curve was linear in the range of 49.5～990 ng(r=0.999 9). The detection limits was 89.3 pg, and average recovery was 99.1%, RSD was 0.42%. CONCLUSIONS: This method is rapid, accurate sensitive and can be used for the quality control of pirenoxinum natricum eye drops.

OBJECTIVE: To establish the HPLC method for the content determination of baicalin in Xiaoer zhike granule. METHODS: YWG C18 (150 mm?4.6 mm,10 ?m) column was used and the mobile phase consisted of methnol-water-phosphoric acid (47 ∶ 53 ∶ 0.2) with the flow rate of 2.5 mL?min-1. The detection wavelength was set at 280 nm and injection volume was 10 ?L. RESULTS: The linear range of baicalin was 25～200 ?g?mL-1(r=0.999 6) with an average recovery of 99.57%(RSD=1.9%,n=6). CONCLUSION: This method is sensitive, simple and accurate for the quality control of Xiaoer zhike granule.

Objective To investigate the expression of high mobility group box 1 (HMGBI) of TLR4 endogenous ligand and distant organ tissue injury after intestine ischemia/reperfusion and drainage of lymph fluid in rats. Methods Twenty-four Sprague-Dawley (SD) male rats (SPF grade) were evenly divided into 3 groups:Sham surgery group,intestine ischemia-reperfusion (I/R) group,and intestine ischemia-reperfusion with drainage of intestine lymph fluid (IR + drainage) group.The injury of distant organs such as lungs,liver,kidney was evaluated;The expression of high mobility group box 1 (HMGBI) of TLR4 endogenous ligand in intestine,lung and liver after the ischemia-reperfusion injury was measured by immunohistochemistry.Result HE stained sections,as well as HMGB1 immunohistochemistry results showed that the injury of ischemia/reperfusion (I/R) group and ischemia/reperfusion (I/R) + drainage group were more severe than that in the sham group.A large number of cells stained in I/R group,indicating that HMGB1 expression increased.The injury in I/R + drainage group was significantly less severe than I/R group.Western blot tests showed that the expression of HMGB1 in jejunum,ileum,liver,lung increased significantly in I/R group after L/R injury.Gray-scale values of HMGB1/β-actin were 0.3145 ± 0.0549、 1.7352 ± 0.3280、1.4443 ± 0.0926、3.1382 ± 0.4202.Lymph drainage significantly alleviated the damage,the expression of HMGB1 were significantly lower (P ＜0.05).Gray-scale values of HMGB1/β-actin were 0.1745 ± 0.0327、 1.1083 ± 0.2098、 1.1862 ± 0.1221、2.1095 ± 0.1993. Conclusion Increased expression of HMGB1 of TLR4 endogenous ligand is associated with intestinal and distant tissue injury during intestinal ischemia-reperfusion injury.Drainage of lymph fluid can block the gutlymph pathway and thus reduce the source of HMGB1 from the intestinal as well as the injury of distant tissue.

Objective To investigate the expressions of Toll-like receptor 4 (TLR4) and high mobility group box 1 (HMGB1) expression on distant tissue during the intestinal ischemia/reperfusion and the effects of ω-3 polyunsaturated fatty acids (ω-3 PUFAs) intervention in rats.Methods Forty-eight Sprague-Dawley male rats,weighing (281.50 ± 22.68) g,were randomly divided into three groups (n =16) after gastrostomy:normal diet (N) group,enteral nutrition (EN) group and EN plus ω-3 PUFAs (PUFA) group.Each group was further divided into lymph drainage (I/R + D) and non-drainage (I/R) sub-groups (n =8 each) according to whether treated with intestinal lymph drainage.All the rats were subjected to 60 min ischemia by clamping the superior mesenteric artery,followed by 120 min reperfusion,while the rats in the I/R + D subgroups were treated with intestinal lymph drainage for 180 min at the same time.Results The interleukin-6 level in lymph in N (I/R + D) group was significantly higher than in the EN (I/R + D) and PUFA (I/R + D) groups (PUFA vs EN vs N:(154.57 ±69.30) ng/L vs (97.58 ±40.34) ng/L vs (85.35 ±23.93) ng/L,P =0.021).Besides,the serum level of HMGB1 in PUFA (I/R + D) group was significantly lower compared to the other 5 groups [PUFA (I/R) vs EN (I/R) vs N (I/R) vs PUFA (I/R + D) vs EN (I/R + D) vs N (I/R + D):(2.95 ± 1.17) μg/L vs (3.86 ±0.99) μg/L vs (4.45 ± 1.73) μg/L vs (1.71 ±1.41) μg/Lvs (2.11±0.56) μg/Lvs (3.13 ±0.79) μg/L,P=0.000],and it also decreased in the PUFA (I/R) and EN (I/R) groups than the N (I/R) group (respectively,P ＜ 0.05).Furthermore,the serum endotoxin level in PUFA (I/R) group was significantly lower compared to the N (I/R) and EN (I/ R) groups[PUFA(I/R) vsPUFA (I/R+D) vsEN (I/R) vs N (I/R):(0.020±0.004) EU/mlvs (0.028 ±0.006) EU/ml vs (0.028 ±0.005) EU/ml vs (0.018 ±0.006) EU/ml,P=0.014].Together the serum tumor necrosis factor-α level in both PUFA (I/R) and PUFA (I/R + D) groups were significantly lower than theEN (I/R),N (I/R) and N (I/R+D) groups [PUFA (I/R+D) vs PUFA (I/R) vs EN (I/R) vsN (I/R) vs N (I/R+D):(12.03 ±6.57) ng/L vs (14.32 ±6.11) ng/Lvs (23.27 ±15.60)ng/L vs (27.42 ± 10.37) ng/L vs (26.87 ± 5.30) ng/L,P =0.013].The jejunum and ileum mucosa in all the I/R groups showed swelling and atrophy and appeared fragile,while the PUFA groups showed less yellow staining and injury than the other two groups (P ＜ 0.05,respectively).In addition,the expressions of TLR4 mRNA in jejunum,ileum,and liver in all the drainage groups were respectively lower than the corresponding non-drainage groups [jejunum:PUFA (I/R) vs EN (I/R) vs N (I/R) vs PUFA (I/R+D) vs EN (I/R+D) vsN (I/R+D):2.32±0.62vs3.08±1.29vs3.50±2.44vs 1.62±0.79vs 1.67±1.11 vs 1.94±0.81,P=0.025; ileum:PUFA (1/R) vsEN (1/R) vsN (1/R) vs PUFA (1/R+D) vsEN (1/R+D) vs N (1/R+D):2.67±1.08 vs 5.22 ± 3.96 vs 6.95 ±4.92 vs 1.70±0.68 vs 1.80±0.29 vs3.68±1.47,P=0.012; liver:PUFA (1/R)vsEN (1/R)vsN (1/R)vs PUFA (1/R+D)vsEN (1/R+D)vsN (1/R+D):5.67 ±1.94 vs 7.50 ±3.89 vs 7.18 ±4.55 vs 1.70 ±0.86 vs 3.90 ± 1.95 vs 4.12 ±2.11,P =0.001],which was consistent with the reduction of HMGB1 and the decrease of nuclear factor-κB activity in intestine,liver,and lung (P =0.000).Conclusions Lymph drainage and ω-3 PUFAs intervention can reduce the production of HMGB1 and inflammation factors,inhibit the expression of HMGB1 and TLR4 mRNA,and thus alleviate distant tissue injury caused by intestinal L/R.

An isatin derivative ofβ-cyclodextrin Schiff base was first synthesized by the condensation reaction between the carbonyl group of isatin and ethylenediamino group of the mono-substituted β-cyclodextrin. The Schiff base ligand was chemically linked to homemade ordered mesoporous SBA-15 silica gel via 3-( triethoxysilyl) propyl isocyanate coupling agent. In this way, a novel isatin derivative of β-cyclodextrin-bonded SBA-15 stationary phase ( ISCDP) was prepared for HPLC. Its chemical and physical parameters were characterized by infrared spectroscopy, mass spectroscopy, elemental analysis, thermogravimetric analysis, scanning electron microscopy, transmission electron microscopy, X-ray diffraction and BET specific surface area analysis. The basic chromatographic property of ISCDP was evaluated by using polar halogenated uracils and disubstituted benzene positional isomers as solute probes in reversed-phase chromatography. ISCDP was also used to enantioseparate twoβ-blocker drugs in polar organic mode and two dansyl amino acids in reversed-phase mode, respectively. The related chromatographic separation mechanism was also discussed. Above studies were expected to provide experimental basis for the practical application of ISCDP in the future. The results showed that the introduction of isatin indole ring could enhanced the reversed-phase chromatographic separation ability of ISCDP for halogenated uracils within 7 min. The new packing also exhibited high stereoselectivity for the position isomers of nitroaniline, aminophenol and benzenediol, in which the para isomers were finally eluted due to strong inclusion interaction between the isatin derivative of β-cyclodextrin ligand and the isomers. Meanwhile, the introduction of isatin indole ring could also improve the chiral separation ability of ISCDP. For example the fast enantioseparations of chiral β-adrenergic blockers and dansyl-amino acids on ISCDP were achieved within 20 min (Rs>1. 3). Obviously, besides hydrophobicity, various synergistic interactions could enhance the separation selectivities of the new stationary phase for chiral and achiral analytes, including dipole-dipole, hydrogen bonding,π-π and inclusion interactions. The ordered pore structure of SBA-15 facilitated to fast and efficient separation and analysis for drugs with good permeability and low mass transfer resistance.

Objective: To assess the potential significance of proliferating cell nuclear antigen (PCNA) labelling indexes (PCNA-LI) and AgNORs number in evaluation of proliferative activity of Wilms′ tumor. Methods: A silver staining for AgNORs and an immunohistochemical method PCNA staining were performed on the biopsy specimens taken from 34 children with Wilms′ tumor. Results: No significant differences were observed between PCNA-LI and the pathological types and clinical stages, whereas fraction of S-phase and PI and the number of AgNORs were significantly higher in patients with PCNA-LI≥25% than that in patients with PCNA-LI<25%. The number of AgNORs/cell correlated well with both pathological types and clinical stages. The combination of PCNA-LI and AgNORs can accurately reflect the proliferative activity of cancer cells in Wilms′ tumor. Conclusion: The current pathological types and clinical stages may reflect the aggressive activity in Wilms′ tumor, but insufficient. The simultaneous determination of PCNA-LI and AgNORs count could be used as the essential complementarity of conventional pathological types and stages for accurate evaluation of biologic behaviour of Wilms′ tumor.

A 6-azido-β-cyclodextrin was synthesized and derivatized with p-nitrophenyl isocyanate as chiral ligand. Following that the ligand was chemically bonded to mesoporous SBA-15 via a ‘Click Chemistry ’ reaction of the azido group with alkynyl group. A new p-nitrophenylcarbamoylatedβ-cyclodextrin bonded SBA-15 chiral stationary phase ( NPCSP ) for HPLC was obtained. The new stationary phase was first used to enantioseparate propranolol in human plasma under the polar organic solvent mode. The effects of methanol content , additive concentration of glacial acetic acid/triethylamine in mobile phase and the temperature on the enantioseparation were studied. The optimal chromatographic conditions were as follows: mobile phase was acetonitrile/methanol/glacial acetic acid/triethylamine (90:10:1. 25:2. 25, V/V), temperature 288 K, flow rate of 0. 5 mL/min, injection volume of 20 μL, detection wavelength at 290 nm. The resolution was 2. 04 with a short run time (< 15 min) under the above conditions. The composition of propranolol in plasma was quantitatively measured by HPLC-MS selected ion monitoring mode ( [ M +H ]+ m/z 260 . 10 ) with hydrochlorothiazide as internal standard. And linear range was 2. 5-250 μg/L and with a good linear relationship. The detection limit was 1 μg/L according to S/N=3. The experimental results showed that the chiral stationary phase exhibited excellent chiral separation ability to propranolol and the analysis method for propranolol in plasma was sensitive, accurate, simple and fast, which could be used for the determination of propranolol in plasma and pharmacokinetic studies.