10.3969/j.issn.1000-8179.2013.12.016

OBJECTIVETo compare the effects between moxibustion at Guanyuan (CV 4), Shenshu (BL 23), Zusanli (ST 36) and western medication on immune function in children with cerebral palsy.

METHODSA total of 230 children with cerebral palsy were randomly divided into an observation group and a control group, 115 cases in each one. Patients in the observation group were treated with warm moxibustion at Guanyuan (CV 4), Shenshu (BL 23) and Zusanli (ST 36). Patients in the control group were treated with oral administration of pidotimod 10 mL every time. The treatment was given once a day, and 30 days were considered as one session for total 90 days. The changes of T-lymphoctyte subgroups, serum immunoglobulin and development quotient were compared 30 days, 60 days and 90 days into treatment respectively; also the occurrence rate.of disease was observed during 6-month and 12-month follow-up visit.

RESULTSThe T-lymphoctyte subgroups (CD3+, CD4+, CD4+/CD8+), serum immunoglobulin (IgG, IgA) and development quotient were significantly improved 30 days, 60 days and 90 days into treatment (P < 0.01, P < 0.05). Regarding the changes of CD3+, CD4+, CD4+/CD4+, IgG, IgA and development quotient, the control group was superior to the observation group 30 days into treatment (all P < 0.05), and the control group was similar to the observation group 60 days into treatment (all P > 0.05), and the observation group was superior to the control group 90 days into treatment (all P < 0.05). There was no significant difference of CD8+ and IgM before and after treatment in two groups (all P > 0.05). The rate of adverse events was 7.0% (8/115) in the observation group, which was lower than 23.5% (27/115) in the control group (P < 0.01); during 6-month and 12-month follow-up visit, the occurrence rate of disease in the observation group was lower than that in the control group (P < 0.05).

CONCLUSIONMoxibustion at Guanyuan (CV 4), Shenshu (BL 23) and Zusanli (ST 36) can improve immune function of children with cerebral palsy, which is superior to pidotimod.

Acupuncture Points ; Cerebral Palsy ; immunology ; therapy ; Child, Preschool ; Female ; Humans ; Immunoglobulin A ; immunology ; Immunoglobulin G ; immunology ; Infant ; Male ; Moxibustion ; T-Lymphocyte Subsets ; immunology ; Treatment Outcome

Aim To explore the role of KATP in the pa-raventricular nucleus in inflammatory pain. MethodsMale Sprague-Dawley rats,250~280 g, were randomly assigned into 5 groups ( each, n =6 ): Normal group, Saline group ( for control, subcutaneous injection of 100 μl of saline into the plantar surface of the left hindpaw) , CFA group ( subcutaneous injection of 100μl of complete freund's adjuvant into the plantar sur-face of the left hindpaw) , Vehicle group ( treated with dimethylsulfoxide), KATP selective agonist group(trea-ted with diaoxide) . Rats in each group were tested for TWL with radiant heat apparatus. Immunofluorescent technique was used to label KATP in PVN and c-Fos in lumber spinal cord. Three days after injected with CFA, a selective KATP agonist, diaoxide, was injected into one side of PVN to test its effect on inflammatory pain and c-Fos expression in lumber spinal cord. Re-sults ① Compared with pre-operation and saline group, rats showed significantly lower TWL on day 1, 3, 7 after injection of CFA;the numbers of KATP posi-tive cells were significantly lower; the numbers of c-Fos positive cells were significantly higher. ② Com-pared with those of vehicle group, TWL and the num-ber of c-Fos in lumber spinal cord were both signifi-cantly lower after injection of diaoxide into one side of PVN. Conclusion KATP in PVN is related to inflam-matory pain.

Objective To establish and optimize AFLP reaction system used in providing necessary technique basis for genetic diversity analysis in Asarum sieboldii.Methods Leaves of A.sieboldii were used as experimental materials to analyze various essential elements of the whole process,such as quality of extracted DNA,time of restriction digest and ligation,concentration of Mg2+,primers and Taq DNA polymerase during PCR amplification etc.,so that the most optimal AFLP reaction system could be built up. Results The optimal AFLP reaction system of A.sieboldii has been constructed: in the genomic DNA extraction,mercaptoethanol being utilized and samples being incubated about 30 min at 65 ℃;the genomic DNA being digested by Trul Ⅰ and Pst Ⅰ for 3 h respectively;in PCR amplification,the final concentration of Mg2+ being 1.5 mmol/L and the volume of Taq DNA polymerase being 0.2 ?L.Conclusion The present reaction system for A.sieboldii is able to gain the favorable results of AFLP analysis and can be used in the genetic diversity research of the species.

Dried herb of Delphinium brunonianum Royle (Ranunculaceae) has long been used under the herbal name "Xiaguobei" (Delphinii Brunoniani Herba) in traditional Tibetan medicine and prescribed for the treatment of influenza, itchy skin rash and snake bites. In order to find a useful and convenient method for the identification of microscopic features, the technique of fluorescence microscopy was applied to authenticate "Xiaguobei" of Tibet. The transverse sections of stem and leaf, as well as the powder of "Xiaguobei" were observed to seek for typical microscopic features by normal light and fluorescence microscopy. A style-like, single-cell glandular hair containing yellow secretions on the leaf, young stem and sepal of "Xiaguobei" was found. Under the fluorescence microscope, the xylem and pericycle fiber group emitted significant fluorescence. This work indicated that fluorescence microscopy could be an useful additional method for the authentication work. Without the traditional dyeing methods, the main microscopic features could be easily found by fluorescence microscopy. The results provided reliable references for the authentication of "Xiaguobei".

Objective:To explore the effect of the synthetic immunomodulator CH 2b with a thiazolidin-4-one ring on the pathogenesis of rheumatoid arthritis (RA) mice induced by glucose-6-phosphate isomerase(GPI) mixed peptides.Methods:hGPI325-339 ,hGPI469-483 peptide fragments were mixed with complete freund′s adjuvant fully ,DBA/1 mice were givien subcutaneous injection of the emulsifiers,pertussis toxin to strengthen immunity.And then RA mice were intervened with α-GalCer and CH2b,the changes of body weight ,ankle joint symptoms scores were observed .The joint tissues stained with hematoxylin and eosin ( HE) was used to evaluate the inflammatory cells.Fluorescence-activated cell sorting ( FACS) was used to detect the frequency changes of iNKT cells .Cytometric bead array(CBA) was used to analyze the levels of serum cytokines TNF-α,IL-6,IL-4,IFN-γ.Results: Compared with the model group,α-GalCer,CH2b could reduce the inflammation of the model mice ,significantly improve the body weight growth and the joint swelling(P<0.05).The inflammatory cells infiltration were decreased.More importantly,CH2b improved the frequency of iNKT cells in peak,the difference was statistically significant (P<0.05).The levels of TNF-α,IL-6,IFN-γin serum were significantly decreased(P<0.05) by CH2b.α-GalCer,CH2b could decrease the levels of TNF-α,IL-6,IFN-γ,and the two groups had no statistically significant difference(P>0.05).Conclusion:Immunomodulator CH2b by activating iNKT cells affect the secretion of inflammatory cytokines ,and it relieved the GPI induced arthritis .

Objective To investigate the effects of a novel synthetic immunostimulator CH2b containing thiazolidin-4-one on the function of invariant nature killer T (iNKT) cells isolated from patients with active rheumatoid arthritis (RA).Methods Peripheral blood mononuclear cells (PBMCs) isolated from patients with active RA were in vitro cultured with α-Galcer and IL-2.The iNKT cells were separated by using magnetic activated cell sorting (MACS) method.The effects of CH2b on the proliferation of iNKT cells were analyzed by using MTT assay.MILLIPLEX MAP Human Cytokine/Chemokine kit was used to measure the levels of IFN-γ and IL-4 in the supernatants of iNKT cell culture.The expressions of IFN-γand IL-4 at mRNA level in iNKT cells were analyzed by RT-PCR.Western blot assay was used to detect the levels of T-bet and GATA-3 in iNKT cells.Results CH2b significantly enhanced the proliferation of IL-2 activated iNKT cells isolated from the patients with active RA.CH2b promoted the secretion of IL-4,resulting in a decrease in the ratio of IFN-γ/IL-4.Moreover,CH2b promoted the expressions of GATA-3 and IL-4 at mRNA level in iNKT cells.Conclusion The novel immunostimulator,CH2b,might enhance the immunoregulatory effects of iNKT cells by promoting the GATA-3 pathway-mediated secretion of Th2-1ike cytokines and inducing the differentiation of Th0 to Th2 cells.

Objective To investigate the perinatal outcome of fetus with increased nuchal translucency (NT) at first trimester.Methods The thickness of NT above 95th percentile of the fetuses with same crown-rump length (CRL) was set as the criteria of increased NT.The outcomes of fetuses with increased NT during early pregnancy from Jan.2008 to Dec.2009 in Guangzhou Women and Children's Medical Center were followed up.The information of ultrasound at second trimester,pregnant complications and delivery outcome were collected.All infants were followed up for 3 months after birth and were divided into four groups according to their different thickness of NT.The relationship between NT thickness and perinatal outcome were analyzed with single factor analysis of variance and multiple comparison method.Results Among the 178 cases we followed up,there were 2 spontaneous fetal losses and 19 terminations whose reasons were Down syndrome (n=6),severe a thalassemia (n =5),fetal malformations (n =7) and social factor (n =1).Among the 157newborns delivered,one was found with congenital heart disease.The rate of abnormal infants was 11.8％ (21/178) and the detection rate of abnormal infants was 9 5.2％ (20/21).Healthy living rate of fetus with NT thickness between 95th percentile and 2.9 mm was 96.1％ (122/127); 82.4％ (28/34) for those with NT thickness between 3.0 mm and 3.9 mm; and 35.3％ (6/17) when NT≥ 4.0 mm.Conclusions Increased NT might have close relationship with poor pregnant outcome.The thicker the NT,the lower the healthy living rate of the fetus.The pregnant outcome is very poor if NT≤4.0 mm.

Objective To explore the effects of C-pseudonucleosides bearing thiazolidin-4-one as immunostimulants on differentiation and activation of human lymphocytes. Methods Peripheral blood mononuclear cells (PBMC) were isolated from healthy adults,and then incubated with immunostimulants (CH1a,CH2a,CH1b,CH2b and pidotimod).After 48 h,we collected the supernatants and then detected the concentrations of IL-2,IL-4 and IFN-γ using ELISA.After 72 h,the proliferation was detected using MTT method.PBMC incubated with immunostimulants (CH1a,CH2a,CH1b,CH2b and pidotimod),after 72 h,the cultural cells were collected and CD expressions of lymphocytes were analyzed by flow cytometry.Results All samples could stimulate proliferation of T cells.Immunostimulants CH1a,CH2a and pidotimod could elevate the expressions of CD3,CD4,CD19 and CD16CD56,and stimulate the secretions of IL-2 and IFN-γ. Immunostimulants CH1b and CH2b could elevate the expressions of CD3,CD4,CD19 and CD16CD56,and stimulate the secretions of IL-2 and IL-4. Conclusion Immunostimulants CH1a and CH2a could differentiate Th0 into Th1 and promote the proliferation of B cells as well as NK cells.However,immunostimulants CH1b and CH2b could differentiate Th0 into Th2 and promote the proliferation of B cells and NK cells.

[ ABSTRACT] AIM:To establish an animal model of rheumatoid arthritis ( RA) in DBA/1 mice induced by im-munodominant mixed peptides derived from glucose-6-phosphate isomerase (GPI).METHODS: The DBA/1 mice were immunized with emulsified mixed peptide fragments of hGPI 325-339+hGPI469-483 or single peptide hGPI325-339 in com-plete Freund′s adjuvant by subcutaneous injection to induce the model of RA .Body weight , ankle joint symptom scores , the pathological change of the ankle joint , the levels of CD4 +T cells in the spleen and peripheral blood , the proportion of iNKT cells in the peripheral blood , and the levels of TNF-αand IL-6 in serum were detected to evaluate and analyze the model.RESULTS:The hind paw of the model mice appeared red swelling on the 8th day, and then aggravated gradually to the limbs.The red swelling reached peak on the 14th day, and then relieved gradually .Inflammation response dominated by lymphocytes and monocytes was observed in the ankle joint .The inflammatory effect of mixed peptides was more obvious than that of the single one (P<0.05).Compared with control group and the mice treated with single peptide , the weight gain was slow, the amount of CD4 +T cells in the peripheral blood and spleen were increased , the proportion of peripheral iNKT cells in the inflammatory peak was decreased (P<0.05), and the serum level of TNF-αwas increased significantly ( P<0.05) in the mice treated with mixed peptide fragments .CONCLUSION: The immunological characteristics of RA model induced by mixed GPI peptides in DBA/1 mice is closer to that in RA patients , especially in the immunopathology of iNKT cells.Therefore, this model can be used as a new tool for studying the mechanism and immunological intervention of RA.