[Effect of moxibustion on acupoints at "opening" time on telomere length and expression of liver cell cycle regulators in aging rats].

OBJECTIVE: To observe the effects of "lingguibafa" moxibustion performing at the appropriate acupoints at their "opening" time on telomere length，expressions of p53 of tumor supressor genes and retinoblastoma gene(Rb)in the liver of aging rats，so as to explore its mechanisms underlying delaying senescence. METHODS: Forty male SD rats were randomly divi-ded into normal, model, prevention and treatment groups, with 10 rats in each group. The rat model was established by intrape-ritoneally injection of D-galactose (200 mg/kg) once a day for 42 days. The rats in the prevention group were given "lingguibafa" moxibustion during modeling, while those in the treatment group were given "lingguibafa" moxibustion after modeling. Quantitative real-time PCR was used to detect the telomere length and the mRNA expressions of p53 and Rb，ELISA was used to detect the protein contents of p53 and Rb in the liver tissues. RESULTS: Compared with the normal group, the relative telomere length of the model group was significantly shortened (P<0.01), the mRNA expressions and protein contents of p53 and Rb were significantly increased (P<0.01). After intervention and in comparison with the model group, the relative telomere length of the prevention group and the treatments group were significantly increased (P<0.01), and the expressions of p53 and Rb mRNA and protein contents were significantly reduced (P<0.01, P<0.05). There were no significant difference between the prevention and the treatment groups in the abovementioned indexes (P>0.05). CONCLUSION: Moxibustion on acupoints at "opening" time can inhibit the shortening of telomere length and the down-regulation of the expressions of p53 and Rb in aging rats, which may contribute to its function in delaying the process of cell senescence.

[Effect of moxibustion at "acupoint-open on-time" on immune function and nuclear factor-κB signaling pathway in aging rats].

OBJECTIVE: To observe the effect of moxibustion at "acupoint-open on-time" （Linggui Bafa） on immune function and nuclear factor-κB (NF-κB) signaling pathway in aging rats, so as to reveal its underlying mechanism of anti-agingt. METHODS: Forty SD rats were randomly divided into control, model, "Linggui Bafa" and "Shenque (CV8)" groups (n=10 in each group). Aging model was established by intraperitoneal injection of D-galactose (300 mg/kg) for 42 days. Moxibustion was applied at "acupoint-open on-time" or CV8 once daily for 28 days. The serum levels of TNF-α, IgG, IgA and IgM were detected by ELISA. The spleen index of rats was calculated. The expression of NF-κB p65 and NF-κB inhibitor protein (IκB) α mRNA of spleen tissues was detected by real-time PCR. RESULTS: Compared with the control group, the expression level of serum TNF-α, the mRNA expression levels of NF-κB p65 and IκBα in the model group was significantly increased (P<0.01), while the expression levels of serum IgG, IgA and IgM, and the spleen index in the model group were significantly decreased (P<0.01). After the treatment, mode-ling-induced increase of serum TNF-α, NF-κB p65 and IκBα mRNA expression levels and decrease of serum IgG, IgA, IgM levels, the spleen index were all completely reversed in the two treatment groups relevant to the model group (P<0.01, P<0.05). No significant change was found in the above-mentioned indexes between the two treatment groups (P>0.05). CONCLUSION: Moxibustion at both "acupoint-open on-time" and CV8 can enhance the immune function of aging rats, which may be related to its effects in inhibiting NF-κB signaling pathway.

Effect of orthodontic force on inflammatory periodontal tissue remodeling and expression of IL-6 and IL-8 in rats.

OBJECTIVE: To investigate effect of orthodontic force on inflammatory periodontal tissue remodeling and expression of IL-6 and IL-8 in rats. METHODS: Eighty SD rats were randomly divided into 4 groups, blank control group (group A) with 5 rats, treatment normal group (group B) with 25 rats, inflammation control group (group (group C) with 25 rats, inflammation treatment group (group D) with 25 rats. Immunohistochemistry and histomorphometric analysis was performed to measure the expression of IL-6, IL-8 and the first molar to the recent movement in the distance. RESULTS: The expression of IL-8 reached a maximum on day 5 and declined thereafter in group B; the expression of IL-6 reached a maximum on day 5 in group B. The expression of IL-6 and IL-8 was gradually weakened with time in group C. The expression of IL-6 and IL-8 were high, and reached a maximum on day 5 and declined thereafter in group D. AD of positive cells in group D were higher than group B at each time point (P<0.05). The time which 0.49 N orthodontic force was loaded was longer, orthodontic tooth movement distance was greater. Movement distance in group D were longer than group B (P<0.05). CONCLUSIONS: Orthodontic force as well as inflammatory stimulus can evoke the expression of IL-6 and IL-8. Under the combined effects of inflammation and orthodontic force, the expression of IL-6, IL-8 will increase.

[Estimating genetic distance and phylogenetic tree of HPA-1-3, 5, and 15 in different populations].

According to the human platelet alloantigens (HPA) polymorphisms in five systems, the distributions of HPA-1 -3, 5, and 15 systems in 1 000 Chinese donors were carried out by using a polymerase chain reaction with sequence-specific primers (PCR-SSP) method. The genetic distance and phylogenetic tree between Chinese Hans and other populations were estimated by using DISPAN and PHYLIP software. As presented by the phylogenetic tree, Asian had a convergence with European first, and grouped together with African. Beninese which came from Africa was on the top of dendrogram. Indian was located between Asian and European. Brazilian was converged with other Europe populations. Oceanian Polynexiya had been shown specifically to cluster with Asia populations. These results proved the "out of Africa theory" from one side, and it also confirmed that early migration of Asian is from south to southeast, and east Asia., thus it is probable that Europeans are migrated from south to north, and west Europe. As genetic distance was estimated effectively by HPA systems, HPA systems could serve as the genetic marker in human migration and evolution research.

HLA-A, -B, -DR haplotype frequencies from DNA typing data of 26,266 Chinese bone marrow donors.

HLA phenotypes of 26,266 Chinese individuals who were recruited as potential hematopoietic stem cell donors by the Shanghai Red Cross Marrow Donor Registry, part of the China Marrow Donor Program, were determined for HLA-A, -B, and -DRB1 alleles at low to intermediate resolution using DNA-based typing methods. The large sample size of the study allowed accurate calculation of the Chinese HLA haplotype frequencies. The observed alleles correspond to 19 HLA-A, 44 -B, and 13 -DR split antigens. The serologic equivalents of HLA-A36, -A80, -B78, and -DR18 alleles were not observed. A total of 2,241 distinct HLA-A, -B, -DRB1 haplotypes were identified. Three-locus haplotype frequency was estimated using the maximum likelihood method. The lowest haplotype frequency that can be reliably estimated at a 95% confidence level was 0.000057. Using this cutoff value, 1,220 haplotypes (54%) were statistically reliable and their cumulative haplotype frequency was 0.9730. The cumulative haplotype frequency of the remaining 1,021 haplotypes (46%) was 0.0270. A regression equation of p = 0.192 log N - 0.576 was derived to estimate the probability (p) of finding an HLA-A, -B, -DR split antigens-matched donor in a pool of N Chinese donors.

[The polymorphism distributions of MICA and MICB microsatellite in Guangdong Han population].

OBJECTIVE: To investigate the genetic polymorphism of microsatellite in the exon 5 of MICA gene and the intron 1 of MICB gene in Guangdong Han population. METHODS: One hundred and six samples of Guangdong Han population were genotyped by polymerase chain reaction and fluorescent technique (6-FAM). Gene frequency, power of discrimination, expected heterozygosity, polymorphism information content and probability of paternity exclusion were calculated. RESULTS: The genotype distributions of MICA and MICB microsatellite met Hardy-Weinberg equilibrium. MICA A5 was the most common allele (0.2877), whereas A4 was the least popular one (0.1321). The genotype distribution frequencies of A5-5.1 (14.15%) and A5-5 (10.38%) are high. MICB CA14 was the most common allele (0.3255), and CA19,28 was the least popular one (0.0047). CA27 was not observed. The genotype distribution frequency of CA14-CA14(14.15%) is high. CONCLUSION: The microsatellite of the exon 5 of MICA gene and the intron 1 of MICB gene could be used as the genetic markers of Chinese population in the studies of anthropology, linkage analysis of genetic disease genes, individual identification and paternity test in forensic medicine.

[The study on the haplotype of MICA and MICB microsatellite locus in Guangzhou Han population].

This study is to investigate genetic polymorphisms and haplotypes of microsatellite locus in the exon 5 of the MICA gene and intron 1 of the MICB gene based on 106 samples of Guangzhou Han Population by polymerase chain reaction and fluorescent technique (6-FAM). The corresponding haplotype frequencies, linkage disequilibria values and relative linkage disequilibria values were estimated based on population data. The results show that the genotype distributions of MICA and MICB microsatellite meet Hardy-Weinberg equilibrium in Guangdong Han population. In total, 5 alleles of MICA microsatellite locus and 14 alleles of MICB microsatellite locus were observed. MICA A5 was the most common allele (0.2877), whereas A4 was the least popular one (0.1321). MICB CA14 was the most common allele (0.3255), and CA19 and CA28 were the least popular ones (0.0047). CA27 was not observed. Twenty-one kinds of MICA-MICB haplotypes occurred at frequencies of more than 1% (linkage disequilibria value>0). The common MICA-MICB haplotypes were A5-CA14 (16.73%) , A5.1- CA18 (8.75%), A4- CA26(3.76%),A9-CA15(3.66%) and A6-CA21(2.61%) (chi(2)>3.84, P<0.05) , and they were strong linkage disequilibria. The polymorphisms and haplotypes distributions of MICA and MICB microsatellite locus in Guangzhou Han population have their own genetic characteristics. The microsatellite locus of the exon5 of the MICA gene and intron 1 of the MICB gene could be used as the genetic markers in the studies of anthropology, linkage analysis of genetic disease genes, individual identification and paternity test in forensic medicine.

[Study on HLA haplotypes in Jiangsu-Zhejiang-Shanghai Han population].

This study is to investigate HLA haplotypes in Jiangsu-Zhejiang-Shanghai Han population based on 166 families by serological and molecular biological HLA typing methods and to analyze the distribution characteristic of HLA haplotypes. The results showed that allele frequencies of more than 10% for HLA antigens were A2, A11, A24, B13, B46, B60, DRB1 *04, DRB1 *08, DRB1* 09, DRB1 * 12 and DRB1 * 15. In the analysis of HLA haplotypes, 128 kinds of A-B haplotypes and 182 kinds of B-DRB1 haplotypes were found, comprising 19.28% (128/664) and 27.41%(182/664) of total theoretical haplotypes, respectively. 18 kinds of A-B haplotypes and 23 kinds of B-DRB1 haplotypes occurred at frequencies of more than 0.5% (linkage disequilibrium value, delta > 0) .351 kinds of A-B-DRB1 haplotypes were found, comprising 52.86% (351/664) of total theoretical haplotypes, and 8 kinds of A-B-DRB1 haplotypes occurred at frequencies of more than 0.5% (delta > 0). The common A-B-DRB1 haplotypes were A30-B13-DRB1 * 07(4.22%), A2-B46-DRB1 * 09(3.77%), A33-B58-DRB1 * 17(3.01%), A33-B58-DRB1 * 13.1 (1.81%) and A11-B75-DRB1 * 12(1.51%). The HLA haplotype distribution of Jiangsu-Zhejiang-Shanghai Han population has its own genetic characteristics, so it suggests this population is between southern and northern Han population. The HLA polymorphism of Chinese Han population is more abundant in East Asian populations.

[The genetic characteristic of HLA-DRB1 locus in the Jiangsu-Zhejiang-Shanghai Han population and a comparison of its frequency distribution with that of other populations].

OBJECTIVE: To investigate the genetic polymorphism of HLA-DRB1 locus in Jiangsu-Zhejiang-Shanghai Han population and analyze the characteristic of the allele frequency distribution in comparison with that of other populations. METHODS: The technique of polymerase chain reaction-sequence specific primers (PCR-SSP) and reverse polymerase chain reaction-sequence specific oligonucleotide probe (PCR-SSOP) was adopted in genotyping a sample of 626 unrelated healthy individuals collected from a Chinese Han population in Jiangsu-Zhejiang-Shanghai area. HLA-DRB1*0101-1001, DRB3, DRB4 and DRB5 were detected. The allele frequency of HLA-DRB1 was calculated, and the allele frequency distribution of HLA-DRB1 in this population was compared with the results from other populations. RESULTS: HLA-DRB1*0101, 0301, 0701, 09012, 1001, 1201, 1202, 1301/02, 1303/04, 1401/04/05, 1402/03/1305, 1501/02, 16021 and 04xx, 08xx were detected in Jiangsu-Zhejiang-Shanghai Han population. The common HLA-DRB1*allele included 09012(17.97%), 04xx(12.53%), 1202(11.42%) and 1501/02(11.02%). The polymorphism information content is 0.9024, and expected heterozygosity is 0.9634 in Jiangsu-Zhejiang-Shanghai Han population. CONCLUSION: The HLA-DRB1 distribution of Jiangsu-Zhejiang-Shanghai Han population shares some genetic characteristic with other Han populations, but it exhibits its own characteristic, suggesting the intermediate state of this population between the southern and northern Han populations. The polymorphism of HLA-DRB1 of Jiangsu-Zhejiang-Shanghai Han population is the most abundant one in this study.