BACKGROUND: Polymorphism of exon 5 +3953C/T of interleukin-1β (IL-1β) gene is found in some studies. This gene polymorphism is related to stomach cancer, rheumatism, arthritis and other diseases. However, what is the distribution property of genotype of IL-1β gene in Guangxi Zhuang population? OBJECTIVE: To observe the distribution of the polymorphism of exon 5 +3953C/T of 1L-1β gene in Chinese Guangxi Zhuang population and Han population, and compare its distribution with Caucasians and African white people.DESIGN: Contrast observation.SETTING: Department of Histology and Embryology,Youjiang Medical College for Nationalities.PARTICIPANTS: Persons who received the health examination in the Affiliated Hospital of Youjiang Medical College for Nationalities were recruited in the experiment. Among them, 155 were Zhuang nationality, including 80 male and 75 female, aged (37) years, and 195 were Han na tionality, including 100 male and 95 female, aged (38) years. Blood relationship was not found among them,and informed consents were obtained from all the participants.METHODS: This experiment was carried out in the Experimental Center for Medical Sciences,Guangxi Medical University from June 2003 to May 2005. Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) technique was used to detect the polymorphism of exon 5 +3953C/T of IL-1β gene in Guangxi Zhuang population (n=155) and Han population (n=195).Genotype and allele frequency of IL-1β gene of healthy population were directly calculated with counting method, and they were compared between Caucasians and African white people combining with literatures.MAIN OUTCOME MEASURES: Comparison of genotype frequency and allele taking frequency of IL-1β gene (+3953) among different ethnic populations.RESULTS: Subjects who received healthy examination including 155 Guangxi Zhuang nationality and 195 Chinese Han nationality participated in result analysis. ①The frequencies of genotypes and allele of IL-1β gene (+3953C/T) were close between Guangxi Zhuang population and Han nationality (Frequency of genotype CC,CT TT and allele C and T of Zhuang population was 94.8％,5.2％,0,97.4％,2.6％,respectively,and that of Han population was 93.3％,6.7％,0,96.7％,3.3％,respectively,P＞0.05).②There were significant differences in genotype distribution and allele taking frequency of IL-1β gene between Caucasians,African white people and Guangxi Zhuang population,Han population (Frequency of genotype CC,CT TT and allele C and T of Caucasians was 52.2％,43.9％,3.9％,74.1％,25.9％,respectively,and that of African white people was 60.4％,33.2％,6.4％,76.4％,23.6％, respectively, P＜0.01).CONCLUSION: The distribution of polymorphism of exon 5 +3953C/T of IL-1β gene is close between Guangxi Zhuang population and Han population,but significant difference exists in comparison with Caucasians and African white people.

Objective To observe the curative effect of puerarin on Alzheimer's disease (AD) in ovariectomized versus non-ovariectomized rat.Methods 100 6-month-old female SD rats were randomly divided into 5 groups:the control group,non ovariectomized AD group,ovariectomized AD group,non-ovariectomized AD treatment group and ovariectomized AD treatment group (n =20each).Rat model of AD was made by treating with combined D-galactose and sodium nitrite for 60days.Treatment group rats were treated with puerarin for 8 weeks.At the end of the experiment,rats learning and memory abilities were tested by water maze,the serum estradiol,progesterone,follicle stimulating hormone and luteinizing hormone were measured,and the activities of cholinesterase (TchE) and superoxide dismutase (SOD),malondialdehyde (MDA) concentration in brain homogenate were determined.Results Compared with the control group,TchE activity and MDA content were increased,and SOD activity was reduced (all P＜0.01),and learning and memory abilities were significantly decreased in the ovariectomized and non-ovariectomized AD groups.Compared with the model groups,the activity of TchE and MDA concentration were reduced,and SOD activity was increased (all P＜0.01),and learning and memory abilities were significantly improved in the two treatment groups.As compared with non ovariectomized AD treatment group,the increase in SOD and TchE activities and decrease in MAD concentration were much more in ovariectomized AD treatment group (P＜0.05),and the improvement in learning and memory abilities was more significant.Conclusions Puerarin has a therapeutic effect on rat model of AD,even better on the ovariectomized rat model of AD.

BACKGROUND: Neural stem cells are a kind of special cells possessing self-renewal and multi-directional differentiation potential. They are ideal carriers for studying the occurence, development and development rule of nervous system and their inherent regulation mechanism of molecular biol ogy and cytology. They have wide applicative prospect in the treatment of nervous system disease and injury. Investigating a set of convenient, effective and useful culture system of neural stem cells cultured in vitro is the prerequisite and basis for application. OBJECTIVE: To observe the effect of serum culture medium containing basic fibroblast growth factor on the in vitro culture of neural stem cells of mice. DESIGN: Single-sample observation. SETTING: Department of Histology and Embryology, Youjiang Nationality Medical College. MATERIALS: Five 2-month-old Kunming white mice of either sex were used in this experiment. METHODS: This experiment was carried out at the Department of Histology and Embryology, Guangxi Medical University from June 2003 to May 2005. ① The cerebral cortical cells were isolated from cerebral cortex and cultured in vitro of the adult mouse cerebral cortex and made into cell suspension , then put in the DMEM/F12 (1:1)culture medium (containing fetal bovine serum of 0.15 volume fraction, basic fibroblast growth factor of 20 μg/L, penicillin of 100 u/mL and streptomycin of 100 u/mL ) .The attachment culture method was used to acquire the clone cells. ② Expression of nestin antigen of clone cells was detected with immunohistochemical technique. MAIN OUTCOME MEASURES: ① Morphological observation of cultured cells. ② Observation of cultured cells stained by haematoxylin and eosin . ③ Identification of neural stem cells. RESULTS: ① Morphological observation of cultured cells: After inoculation, most of the cells attached to the wall within 12 hours, cells were thin and flat. 3 days after inoculation, cells began to grow and proliferate. There were several scores to several hundreds of cell clones on the 5th day after inoculation and cells covered 80%-90% of the bottom of the bottle on the 7th to 8th days. After passage, the cells still grew adhesively with regular cellular morphology and clear borderline. ② Observation of cultured cells stained by haematoxylin and eosin: It was shown that the cells presented round or ellipse. There was little cytoplasm, presenting acidophily. Nucleus was big and round, single in the center, deep stained.③ Identification of neural stem cells: Immunofluorescent detection showed that cell nestin antigen positive. CONCLUSION: After cultured in serum culture medium containing basic fibroblast growth factor, cerebral cortex cells possess very strong reproductive activity and express nestin antigen, still possessing the characteristics of neural stem cells.

BACKGROUND: Based on the multi-directional differentiation potential,the combined application of proper biological materials and growth factor can repair tissue defect.OBJECTIVE: To observe the growth characteristics of rabbit bone marrow-derived mesenchymal stem cells and their osteogenic potential under the induced condition.DESIGN: A single sample experiment.SETTING: The Department of Histology and Embryology of Youjiang Medical College for Nationalities; the Affiliated Hospital of Youjiang Medical College for Nationalities.MATERIALS: This experiment was conducted at the Department of Histology and Embryology of Youjiang Medical College for Nationalities in June 2004. Male new Zealand rabbits, with a body mass of 2.0 to 2.5 kg,were provided.METHODS: Rabbit bone marrow tissue was extracted. After gradient centrifugation, adherent cells were kept for cell passage. Culture medium was used for inducing culture of osteoblast when the passage was stable (50 mL/L of Dulbecco modified culture medium of bovine serum containing 10 mmol/L of β-sodium glycerophosphate, 10 nmol/L of dexamethasone, 50 mg/L of vitamine C). The culture medium was changed every other day. Osteogenic characteristics of the bone marrow-derived mensenchymal stem cells were observed under an inverted microscope after alkaline phosphatase immunohistological staining, osteonectin and osteopontin immunocytochemical staining.MAIN OUTCOME MEASURES: ① Morphological observation of bone marrow-derived mesenchymal stem cells; ② Osteogenic characteristics of bone marrow-derived mesenchymal stem cells.RESULTS: ① Morphological observation of bone marrow-derived mensenchymal stem cells: Primary rabbit bone marrow-derived mesenchymal stem cells could cover the Petri dish fully in 7 or8 days and could passage stably. Generative cells began to passage in 5 or 6 days. ② Osteogenic characteristics of bone marrow-derived mensenchymal stem cells:Alkaline phosphatase immunohistochemical staining showed a lot of brown particles in the kytoplasm, but no colored cells were found in the control group. Osteonectin and osteopontin immunohistochemical detection found that the nucleus was light blue and a lot of brown particles appeared in the kytoplasm, presenting obvious strong positive, but no brown particles appeared in the control group.CONCLUSION: After combined induced culture with dexamethasone, vitamine C and β-sodiumglycerophosphate, rabbit bone marrow mensenchymal stem cells showed the morphological and biological characteristics of osteoblasts with osteoblastic activity, and were able to supply autologous seed cells for constructing in vitro tissue engineered bone in a short period of time.

AIM: To explore the effect of microRNA-146a (miR-146a) on apoptosis of human gastric cancer SGC-7901 cells and the underluing mechanism.METHODS: miR-146a mimic (up-regulated miR-146a expression) and miR-146a inhibitor (down-regulated miR-146a expression) were transfected into the SGC-7901 cells by liposome method.At the same time, miRNA nonsense sequence transfection group as the negative control group (NC group) was set up.RT-qPCR was used to evaluate the levels of miR-146a in the SGC-7901 cells after transfection.The effects of miR-146a on the cell apoptosis and growth were assessed by flow cytometry analysis and CCK-8 assay, respectively.The effect of over-expression or knockdown of miR-146a on transforming growth factor-β-activated kinase 1 (TAK1)/ nuclear factor-kappa B (NF-κB) signaling was evaluated by RT-qPCR and Western blot.RESULTS: miR-146a modulated apoptosis of SGC-7901 cells.Over-expression of miR-146a significantly increased apoptosis, whereas knockdown of miR-146a inhibited the apoptosis of SGC-7901 cells.The expression of TAK1 at mRNA and protein levels was significantly decreased when miR-146a mimic was transfected into the SGC-7901 cells (P<0.05).On the contrast, the expression of TAK1 at mRNA and protein were significantly higher in miR-146a inhibitor transfection group than that in NC group (P<0.05), suggesting that miR-146a negatively regulated TAK1 expression.Moreover, knockdown of TAK1 enhanced the apoptosis of SGC-7901 cells (P<0.01), while over-expression of TAK1 inhibited the apoptosis of SGC-7901 cells(P<0.01).Additionally, both over-expression of miR-146a and knockdown of TAK1 led to a prominent increase in the expression of NF-κB inhibitor protein alpha (IκBα) and a significat decrease in B cell lymphoma-2 (Bcl-2) level in the SGC-7901 cells.CONCLUSION: miR-146a significantly promotes apoptosis of SGC-7901 cells by inhibition of NF-κB pathway via targeting TAK1.

Objective To explore the prevention and treatment of lymphorrhagia due to lymph node dissection in gastric carcinoma. MethodsClinical data of 743 patients undergoing radical gastrectomy plus lymphadenectomy from January 1997 to March 2004 were analyzed retrospectively. ResultsThe patients in D3 or D4 lymphadenectomy suffered from a higher lymphorrhagia rate than those in D2 lymphadenectomy(P

Objective To re-evaluate the immunogenicity of meningococcal serogroups A and C polysaccharide vaccine among a healthy population of age 5 to 59.Methods Pre and post-vaccination ser-um samples were collected from the subjects involved in a randomized , controlled clinical trial conducted in 2005 at Hechi, Guangxi, China.Enzyme-linked immunosorbent assay (ELISA) was performed for the quan-titative detection of specific anti-capsule IgG antibody against meningococcal serogroups A and C in serum samples.Serum bactericidal assay ( SBA) was used to measure the bactericidal antibody activity against se-rougroups A and C bacterial strains .Results Geometric mean concentrations (GMCs) of specific anti-cap-sule IgG antibody were 23.66 μg/ml and 78.83 μg/ml for serogroup A (P<0.05), and 1.23 μg/ml and 51.25 μg/ml for serogroup C (P<0.05) in serum samples of pre and post-vaccination, respectively.Be-fore immunization , 99% of serum samples ( 97/98 ) showed serogroup A-specific IgG concentration ≥2μg/ml, which reached up to 100%(98/98) after vaccination (P>0.05).The percentage of serum sam-ples with serogroup C-specific IgG concentration ≥2 μg/ml rose from 20%to 99% after vaccination ( P<0.05).The ratio of positive serum samples with at least four times of increases in concentration of specific IgG against serogroup A and serogroup C after vaccine immunization were 34% and 100%, respectively . The rSBA geometric mean titers ( GMTs) for serogroups A and C in serum sample of post-vaccination were respectively elevated from 1164 to 5530 (P<0.05), and from 4 to 6225 (P<0.05).The percentage of se-rum samples with rSBA GMTs≥128 increased from 91% to 99% for serogroup A (P>0.05), and from 14%to 96%for serogroup C (P<0.05) after vaccination.The rSBA GMTs with at least four times of in-creases after vaccination were detected respectively in 53% and 100% of serogroups A and C vaccinated subjects.Pearson correlation coefficient between IgG concentration and rSBA GMTs was r=0.15 (pre) and r=0.23 (post) for serogroup A, and r=-0.14 (pre) and r=0.58 (post) for serogroup C (P<0.05). Conclusion This study has demonstrated an efficient and sustained immunogenicity with meningococcal sero -groups A and C polysaccharide vaccine as evidenced by the data from standardized assays of ELISA and SBA .

Objective:To assess the value of spiral CT in diagnosis and treatment of chronic otitis media.Methods:The spiral CT findings of 74 cases including 93 ears proved by operation and pathology were studied.Results:The lesions such as the disruption of the ossicular chain showed in spiral CT or three-dimensional image were in accord with those seen in the operation,the accuracy was 95.7%,the disruption of the ossicular chain and bony erosion in the tympanic cavity and antrum were severe in the typeⅢ chronic otitis media.Conclusion:Spiral CT is helpful to diagnose and definite the chronic otitis media,three-dimensional image can provide valuable information for surgery.

Objective To establish a standardized quantitative enzyme-linked immunosorbent as-say ( ELISA) recommended by WHO for the detection of human IgG antibodies specific for Streptococcus pneumoniae capsular polysaccharides ( Pn PS ELISA ) .Methods According to the WHO recommended standard Pn PS ELISA protocol , capsular polysaccharide concentrations of 13 serotypes (1, 3, 4, 5, 6A, 6B, 7F, 9V, 14, 18C, 19A, 19F, 23F) of Streptococcus pneumonia for coating were optimized;the ELISA plates and AP conjugated goat anti-human IgG antibody for the detection were experimentally selected .Using the established assay parameters assured by testing quality control standards ( QCs) provided by WHO refer-ence laboratory , a panel of 16 LIBP ( Lanzhou Institute of Biological Products Co .Ltd.) QCs were measured for comparison analysis between WHO reference laboratory and LIBP laboratory .In the meantime , the range of IgG concentrations of an internal QC panel 907 for 13 serotypes of pneumococcal capsular polysaccharide were established for routine QC work in LIBP laboratory , and inter-assay precision within LIBP laboratory was assessed as well .Results The standardized Pn PS ELISA assay established in LIBP laboratory met the criteria required by WHO .There was a high correlation between the data collected by WHO reference labora -tory and LIBP laboratory (slope=0.94, coefficient of correlation r=0.97, P<0.05).Eighty one percent of IgG concentrations measured by LIBP laboratory were within the range of ±40%of those measured by WHO reference laboratory , which met the criteria of 75%data falling within the ambit of ±40%of assigned values commonly used for comparison .The ranges of IgG concentration in LIBP QC 907 for 13 serotypes had been established.The inter-assay precision in LIBP laboratory was high with coefficiency of variation ( CV) less than 30%.Conclusion LIBP laboratory has successfully established the standardized ELISA recommended by WHO for quantitative detection of human IgG antibodies against pneumococcal capsular polysaccharides (Pn PS ELISA).The range of IgG concentrations in LIBP QC 907 for 13 serotypes of pneumococcal capsular polysaccharide are also established for routine quality-control practice .

Objective To dynamically analyze the antibodies with regard to in vitro serum bacteri-cidal activity and the quantity of IgG in healthy adults received one dose of immunization with ACYW 135 me-ningococcal polysaccharide vaccine .To investigate the term of protection with polysaccharide vaccine and the correlation between bactericidal titer and IgG concentration .Methods Twenty healthy adults were given one dose of immunization with quadrivalent meningococcal polysaccharide vaccine .Serum samples were col-lected before and after vaccination at specific time points .Test for serum bactericidal activity ( SBA ) and quantitative ELISA were performed to detect bactericidal titers and IgG concentrations in serum samples .Re-sults Certain levels of antibodies against capsular polysaccharides of serogroup A , C, Y and W135 strains had already existed prior to immunization .Moreover, bactericidal titers against serogroup A , Y and W135 strains except serogroup C strain were relatively high .Specific IgG concentrations and bactericidal titers for all serogroup stains were significantly increased on day 15 after vaccination (P<0.05), and then sustained at a high level during a time period from day 30 to day 160.Both IgG concentrations and bactericidal titers dropped to the levels similar to that before preimmunization in about 3 years.No significant correlation was observed between bactericidal titers and IgG concentrations (r<0.3, P>0.05).However, a close correla-tion was demonstrated between GMTs and GMCs of serum samples (r>0.7, P<0.05).Conclusion The geometric mean titers ( GMTs) and geometric mean concentrations ( GMCs) of serum samples collected be-fore and after vaccination at different time points were reliable and consistent parameters for the evaluation of vaccine .The term of protection of quadivalent meningococcal polysaccharide vaccine was about 3 years upon a single dose of immunization .