AIM To study the influence of chronic stress on the level of CNTF and CNTF mRNA in hippocampal neurons of rats. METHODS The chronic stress model was established by chronic unpredictable mild stress, openfield test was performed to detect the behavior of rats. Immunohistochemistry and in situ hybridization were used to observe the level of CNTF and CNTF mRNA. RESULTS Compared to control group, the CNTF like immunoreactivity and signals of CNTF mRNA in situ hybridization in the hippocampal neurons of chronic stress group were significantly decreased. CONCLUSION These results show that chronic stress can significantly decrease the level of CNTF and CNTF mRNA in the hippocampal neurons of rats.

AIM: To identify human leukocyte antigen (HLA)-DRB11454 allele and HLA-DRB1 exon 3 sequence information in the Chinese population, which is significant for organ transplantation, cell transplantation, and human genetics.METHODS: Polymerase chain reaction sequence-based typing (PCR-SBT) was used to identify HLA-DRB1 alleles from 58 donor-recipient individuals who would undergo haemopoietic stem cell transplantation. Medium to high resolution polymerase chain reaction-reverse sequence specific oligonucleotide probe (PCR-RSSOP) was used to identify HLA-DRB1 alleles from 1 268 healthy donors from Guangdong province. The some ambiguous results of HLA-DRB114-associated alleles were confirmed by high resolution polymerase chain reaction-sequence-specific primer typing (PCR-SSP).RESULTS: HLA-DRB11403, 1406, 1410, 1412, 1418, 1425 and 1454 alleles were detected in 1 268 healthy donors.HLA-DRB11454 was confirmed in 8 ambiguous results of HLA-DRB11401/1434/1454 alleles, and HLA-DRB11454 was one of common alleles of HLA-DRB114 allele group in Guangdong population. HLA-DRB114 exon 3 sequence information was confirmed to be polymorphic in Chinese population.CONCLUSION: HLA-DRB11454 and exon 3 of DRB1 are confirmed to be polymorphic in Chinese population, further elucidating that HLA-DRB1 axon 3 sequence information is important for Han population and some minority groups.

Objective To comparatively evaluate the effects of a recombinant Mtb protein ESAT 6-CFP10 ( rESAT6-CFP10 ) and a purified protein derivative ( PPD ) as skin test reagents in guinea pigs . Methods Guinea pigs were sensitized with different Mycobacteria species .After sensitization , all guinea pigs were intradermally injected with rESAT6-CFP10 and PPD.At 48 h after the injection, the size of ery-thema at injection sites was measured by using a double-blind method .For guinea pigs sensitized with viable Mtb, the size of erythema at injection sites were measured at 24 h after the injection .The positive conversion rates of skin test with rESAT 6-CFP10 and PPD were calculated .Results The results of PPD skin test were positive in all guinea pigs sensitized with viable Mtb , killed Mtb and BCG with erythema diameters of (11.4 ±0.9) mm, (11.8±1.1) mm and (13.2±0.8) mm, respectively.Positive skin test with rESAT6-CFP10 was only observed in guinea pigs infected by viable Mtb-showing erythema diameters of (13.7±5.7) mm. The skin test with rESAT6-CFP10 was negative in guinea pigs sensitized by killed Mtb-and vaccinated by BCG.The skin tests by using rESAT6-CFP10 and PPD were performed on randomly selected guinea pigs at ninth day after infection by Mycobacterium tuberculosis H37Rv.At the 2nd week, totally 24 selected guinea pigs showed positive skin test results with rESAT6-CFP10 (24/24) with erythema diameters of (19.9± 3.0) mm, while only 15 out of 24 had positive PPD skin test with erythema diameters of (6.1±5.5) mm. At the 4th week, all guinea pigs showed positive PPD skin test (3/3) with erythema diameters of (12.7± 2.5) mm.Conclusion The skin test by using recombinant ESAT 6-CFP10 protein can effectively distin-guish viable Mtb infection from BCG vaccination and killed Mtb sensitization , which is a more suitable anti-gen than PPD for the early diagnosis of Mtb infection .

AIM: To investigate the survival, migration and differentiation of human bone marrow mesenchymal stem cells (hMSC) in the middle cerebral artery occlusion (MACO) model and to observe the influence on motor function in the animal model. METHODS: hMSC with Hoeschst 33342 were injected into the animal model of MACO after Shenqiye induced for half an hour and their survival, migration, differentiation and the behavior changes in MACO rats were examined. RESULTS: hMSC had good homogeneousness and immunological reaction after implantation. The results showed that hMSC survived in rat brain for a long time over six weeks. As time going, hMSC were found in much more areas in the rat brain. Immunofluorescence staining suggested that implanted hMSC expressed human neuron specific enolase, neurofilament, and glial fibrillary acid protein. At the same time, improvements in abnormal behavior of MACO rats were observed. CONCLUSION: hMSC differentiate into neurons in the brain of rats, which means that hMSC is an ideal seed cells for the therapy of cerebral infarction.

Objective To study the distributive characteristics of HLA-A,B,DRBI alleles and haplotypes patients with ALL in southern Chinese Han.Methods The frequencies of HLA-A,B,DRB1alleles and haplotypes were estimated by Expectation-Maximization method based on the genotypes of 572patients with ALL and 5645 unrelated health donors,and then compared by chi-square test.Results The frequencies of HLA-A33(7.15%vs 9.3%,OR=0.73,P＜0.05),B58(5.93%vs 8.75%,OR=0.64,P＜0.05),DRB1*17(5.15%vs 6.30%,OR=0.82,P＜0.05)alleles and HLA-A33-B58-DRB1*17(2.46%vs 4.14%,OB=0.35,P＜0.05)haplotype were significantly lower in ALL patient groups than that in controls.The frequencies of HLA-A3(2.1%vs 1.26%,OR=1.7,P＜0.05),B51(7.25%vs 5.78%,OR=1.3,P＜0.05)and DRB*12 (16.13%vs 12.99%,OR=1.35,P＜0.05)alleles and A2-B51-DRB1*12(1.24%vs.0.89%,OR=1.66,P＜0.05)haplotype were significantly higher in ALL patient groups than that in controls.Conclusion These results indieated that HLA-A33-B58-DRB1*17 haplotype was a associated with a diminished incidence of ALL.and HLA-A3 auele or A2-B51-DRB1*12 haplotype was weakly associated with ALL.

Objective To establish and evaluate 3 kinds of minimally-invasive valve implantation model in vivo.Methods A novel tissue engineered heart valve(TEHV) manufactured by branched polyethylene glycol cross-linked acellular porcine valve and a minimally-invasive valve implantation system according to the design of Corevalve revalving system were adopted.After anesthesia,18 adult male goats were randomly divided into 3 groups: the ulrasound-directed orthotropic group (group A,n =6),angiography-directed orthotropic group (group B,n =6) and direct-released heterotopic group (group C,n =6),and all received minimally-invasive valve implantation orthotropically or heterotopically.4 weeks later,the valvular function was evaluated by CTA and/or echocardiography.Results All 3 kinds of caprine model were successfully constructed.The operation success rate of each group was A: 66.7％,B: 50.0％ and C: 100.0％,respectively(multiple x2 analysis,group A and B P ＞0.05; group A and C,group B and C,P ＜0.05).The operation-time of each group was A: (79 ± 18) min,B:(61 ±23) min,C: (45 ± 15) min(one-way ANOVA,P ＜0.05).The survival rate at4 weeks was A: 100％,B: 100％ and C: 83.3％ (multiple x2 analysis,P ＞ 0.05).Echocardiography and CTA proved the short-term function of implanted TEHV was satisfactory.Conclusion All 3 kinds of caprine valve implantation model can be established without cardiopulmonary bypass and blood transfusion.The devices and equipments required in group A is relatively simple,but the procedure cost longer time for it is hard to determine the right position by ultrasound.The application of angiography made the positioning much easier in group B while the procedure had to be performed in specific operation room with angiographic apparatus.Group C did rely on neither special equipments nor complex operation,but the valve leaflets cannot work normally,so this model was only suitable for testing in vivo characteristics such as biocompatiblities.

Objective To evaluate the application and the good qualities of high-resolution ultrasound and CDFI-guided mammotome minimally invasive biopsy device in the diagnosis and treatment of breast lesions.Methods The common clinical operations and the lesions which were guided mammotome minimally invasive biopsy device by high-resolution ultrasound and CDFI were contrasted.The effects of treatment were evaluated.Results 307 le- sions of 102 patients were removed by this method,and the operational process was successful.Patients' skin lacera- tions were tiny.Only one lesion was clinically diagnosed as mild blood clot under skin,but without other complica- tions.Conclusion Contrasted with the common clinily operations.the high-resolution ultrasound and CDFI-guided mammotome minimally invasive biopsy device in the diagnosis and treatment of breast lesion is effective,and the scar is tiny.It releases patients' pain.

Objective To establish a guinea pig model of latent Mycobacterium tuberculosis infec-tion for evaluating the effects of therapeutic vaccines .Methods Guinea pigs were subcutaneously inocula-ted with 5.0×103 CFU Mtb.The skin test was performed with 0.5μg recombinant ESAT6-CFP10 protein to detect positive conversion rates at different time points .Two weeks after Mtb inoculation , guinea pigs in model group received 5 mg isoniazid treatment ( three times a week for four weeks ) by oral gavage , while those in control group received normal saline .At the sixth week after Mtb infection , guinea pigs with and without isoniazid treatment were dissected for pathology examination .The pathological scores of liver , spleen and lung, as well as bacteria loads in spleen were compared between two groups .The established guinea pig model of latent infection was then validated by testing two reference vaccines ( AEC/BC02 and AEC/BC03 ) . Results Two weeks after Mtb inoculation , all guinea pigs showed positive EC skin test with induration area of (19.9±3.0) mm.Upon four weeks of isoniazid treatment , the guinea pigs in model group showed no pathological changes with zero scores in the examined organs .No bacterium was detected in spleen of ani-mals from model group.However, the total pathological score was 38.8±16.5 and bacteria load in spleen was (5.1±0.3) Log10 CFU with the guinea pigs from control group .Natural recurrence of tuberculosis in model group was observed after drug withdrawal .The total pathological scores were 48.5±23.9 and 51.3± 23.41.The bacterial loads in spleen were (4.5±1.3) and (4.2±1.1) Log10 CFU and bacterial loads in lung were (4.1±1.2) and (3.4±1.3) Log10 CFU respectively as verified with reference vaccines of AEC /BC02 and AEC/BC03.Conclusion Isoniazid treatment inhibited the proliferation of inoculated Mtb in guinea pigs.A guinea pig model of latent Mycobacterium tuberculosis infection is successfully established with an advantage of good repeatability .Therefore, it can be used to evaluate the effects of therapeutic vaccines on latent Mycobacterium tuberculosis infection.

Objective To establish a suitable guinea pig model for evaluating the protective effects of new TB vaccines in BCG prime-boost regimen .Methods Two different immunization strategies by using the recombinant TB vaccine were employed to boost BCG primed guinea pigs in this study .One was for short-term evaluation with 14 weeks interval between prime and boost immunization and another was for long -term evaluation with 54 weeks interval .In the short-term evaluation group , guinea pigs were boosted twice with the recombinant TB vaccine ( AEC/BC02 ) in every two weeks , while guinea pigs in the long-term evaluation group were boosted for three times with two weeks interval between each injection .A negative con-trol group ( NS→NS) and a BCG control group ( BCG→NS) were both set up in two evaluation groups .One week after the last immunization , all guinea pigs were challenged with M.tuberculosis.Six to seven weeks after bacteria challenge , all animals were euthanized and dissected to evaluate lesion scores of liver , spleen and lung, as well as the viable bacterial load in spleen .Results In the short-term evaluation group , the le-sion scores in those boosted with vaccine (3.33±5.00) was lower than that of BCG control group (5.56± 7.27) (P>0.05) and negative control group (47.00±28.11) (P=0.0001).The difference between BCG control group and negative control group in lesion score was also significant .The animals in vaccine boosted group had lower bacterial loads (0.78±1.55 log10 ) in spleen than that in BCG control group (1.06±1.87) (P>0.05) and negative control group (5.47±0.61) (P=0.0003).In the long-term evaluation group, the lesion score in those boosted with vaccine was lower (5.0±7.6) than that in BCG control group (14.4± 13.5) (P=0.0394) and negative control group (56.9±14.1) (P<0.0001).The animals in vaccine boos-ted group (1.00±1.86 log10) had lower bacterial loads in spleen than that in BCG control group (1.46± 1.94) (P>0.05) and negative control group (5.43±0.56) (P=0.01).There was a significant difference in bacterial load between BCG control group and negative group (P=0.0089).Conclusion The results suggest that the interval time between BCG-prime and boost immunization should be properly prolonged in the guinea pig model used for evaluating the protective effects of new TB vaccines in BCG prime -boost regimen .

OBJECTIVETo study the expression of gastrin(GAS) and gastrin releasing peptide(GRP) in patients with gastric cancer and investigate the clinical significance.

METHODSThe expression of GAS and GRP in sixty patients with gastric cancer was detected by using tissue chip technique and immunohistochemical methods.

RESULTSThe positive rates of GAS and GRP were 30.0% and 11.7% respectively in 60 cases with gastric cancer. The positive rates of GAS and GRP were higher in moderately and poorly differentiated cancers than those in well differentiated cancer (P< 0.05). The positive rates of GAS and GRP were significantly higher in mucinous adenocarcinoma and signet-ring cell carcinoma than those in other types of gastric cancer (P< 0.05). The positive expression of GAS and GRP in gastric cancer was correlated with lymph node metastasis (P< 0.05).

CONCLUSIONTissue chip technique is a feasible,rapid,economic and accurate approach for screening clinical tissue specimens on a large scale.

Adult ; Aged ; Female ; Gastrin-Releasing Peptide ; metabolism ; Gastrins ; metabolism ; Humans ; Immunohistochemistry ; methods ; Male ; Middle Aged ; Neoplasm Invasiveness ; Neoplasm Staging ; Protein Array Analysis ; Stomach Neoplasms ; metabolism ; pathology