Objective To study the relationship between serum amyloid A(SAA)and the common carotid intima-media thickness(IMT)in type 2 diabetic(T_2DM)patients.Methods Sixty-nine patients with type 2 diabetes,and 20 healthy subjects regarded as the normal controls(NC)were enrolled in the study from January to July of 2005.SAA levels were measured using ELISA.The carotid IMT were examined by hypersensitive color Doppler ultrasonography.Results SAA level was significantly elevated in type 2 diabetes group compared with that in the control3.08(2.1~5.06)mg/L vs 1.37(1.07~1.86)mg/L,P

ObjectiveTo investigate changes of sex hormone levels and sexual function in male patients after liver transplantation.MethodsBlood samples were taken from 69 male patients receiving liver transplantation pretransplantation and postoperative 3rd,7th,14th day,1st and 3rd month respectively.Serum estradiol and testosterone were measured by radioimmunoassay.Serum sex hormone binding globulin was measured by ELISA.Sixty-nine discharged adult male liver transplant recipients ( age ranging 24 -45 years) with a normally functioning allograft for at least 6 months were questioned on their pre-and post-operativesexualfunction compared with 23adulthealthy controls.ResultsPretransplantation serum estradiol and sex hormone binding globulin were significantly higher (87.56±31.21 vs.26.00 ±9.12,u=9.30,P＜0.0001,134.50±30.68 vs.51.04 ±12.05,u=12.69,P ＜ 0.0001,respectively),and testosterone was lower than normal controls (2.02 ± 1.28 vs.4.82 ± 1.48,u =-8.73,P ＜0.0001 ) and levels were correlated with MELD scores (r =0.80,P ＜0.0001,r =-0.77,P ＜0.0001,r =0.72,P ＜0.0001,respectively).Sex hormone levels were back to normal on postoperative 2nd week (serum estradiol) or 1st month (serum testosterone and sex hormone binding globulin).Compared with pretransplant status,sexual function improved significantly when evaluated on postoperative 6th month.ConclusionsLevels of serum sex hormone were correlated with MELD scores before liver transplant.After liver transplant,estradiol was back to normal level on postoperative 2nd week,testosterone and sex hormone binding globulin back to normal level on postoperative 1 st month.Sexual function significantly improves after liver transplantation.

AlM:To determine the group distribution characteristics of lachrymal duct obstruction diseases in major Li Miao minority areas of Hainan province.
METHODS: Totally 5 353 residents were selected and researched by randomized cluster sampling in the major Li Miao minority areas of Hainan province. Ocular examination and lachrymal duct flushing were carried out, and questionnaire survey on lachrymal duct obstruction was conducted. The ratio of lachrymal duct obstruction diseases and group distribution characteristics were analyzed based on above research.
RESULTS: The prevalence ratios of lachrymal duct obstruction was 4. 47% in major Li Miao minority areas of Hainan province, with 2. 62% in urban area, and 5. 93% in rural area respectively. Prevalence ratios of men and women group were 1. 69% and 6. 39% correspondingly. Difference between the two groups was statistically significant (χ2 = 67. 2821, P = 0. 0000 ). The highest prevalence ratio was 40-69 year-old group, second one was 70-79 year-old group, especially for women in these groups. The prevalence ratios of Ledong, Lingshui, Baisha and Changjiang county were higher than those of Baoting, Qiongzhong county and Wuzhishan city. No significant difference was found between both eyes.
CONCLUSlON: ln major Li Miao minority areas of Hainan province, lachrymal duct obstruction mainly occurs in 40-79 year-old patients, with specially higher ratio of women. Statistically, significant difference of the prevalence ratio between urban and rural areas exists. The higher prevalence ratio is attributed to age, gender, geographical location, climate condition, health environment and so on. The prevalence ratio is higher in the dry and windy areas than in the humid and less windy areas.

Objective To screen human anti-hypoxia-inducible factor (HIF)-1? scFv of lung adenocarcinoma from large phage antibody library and identify the positive clones. Methods Panning of large phage antibody library against lung adenocarcinoma cell line A549 and HIF-1? was conducted respectively to select specific antibodies. E. coli HB2151 was infected to induce the expression of soluble scFv. The binding activity and specificity were tested by ELISA and immunocytochemical assay. The expression and relative molecular weight of the soluble scFv was detected by SDS-PAGE and Western blot analysis. Results After panning,the target scFv was enriched,and ELISA results showed that positive reactions to HIF-1? were detected in 5 of 10 random clones with a positive ratio of 50%. Immunocytochemical analysis showed the specific affinity of the antibodies to A549 cells. The soluble human anti-HIF-1? scFv fragments of lung adenocarcinoma were expressed in E. coli HB2151 and then confirmed by SDS-PAGE. The result of Western blotting showed that the relative molecular weight of the soluble scFv was about 30?103. The binding activity and specificity were confirmed by ELISA. Conclusion Human anti-HIF-1? scFv of lung adenocarcinoma is successfully obtained with large phage antibody library technique.

Objective To explore the effect of DR on the expression of inhibitory neurotransmitter ,γ-aminobutyric acid ( GABA) and brain derived neurotrophic factor ( BDNF) in the primary visual cortex of young cats .Methods Totally 6 cats were wsed in this study . Nissl staining was used for cortical layer identification and cell counting . Immunohistochemical techniques were utilized to label GABA-and BDNF-positive neurons .Sections were observed under an Olympus light microscope and photographed with a digital camera .The cell density and absorbance of immunoreactivity were measured with Image-Pro Express 6.0 softwares .Results Our results showed that the mean density of Nissl-stained neurons in the primary visual cortex ( V1) of the DR group showed no significant difference from that of the control group . However , the mean density of GABA-immunoreactive neurons in each cortical layer of V 1 in DR cats was significantly higher than that in control cats .The mean immunoreactive intensity of GABA-positive neurons , as indicated by the average absorbance , increased significantly in DR cats relative to control ones . Accompanied with the elevation of GABA expression, DR enhanced BDNF expression in V1, as indicated by an increased mean density of BDNF-positive neurons and BDNF-immunoreactive average absorbance in DR cats relative to controls .Conclusion These results indicate that dietary restriction leads to a concurrent expression upregulation of GABA and BDNF , which may likely compensate for brain functional degradation during senescence and thus delay aging process .

Objective To study CT features of liver abscesses caused by the fasciola hepatica infection , and discuss its pathologic basis.Methods CT images of 15 Patients were collected. All patients underwent both unenhanced and biphasic enhanced CT scanning, then its CT performances were analyzed. Results round and nodular lesions were observed in 15 cases, branching and stripping lesions like dilated bile duct in 9 cases. The density of lesions was inhomogeneous, and the lesions were multifocal and multiform. The liver abscesses caused by the fasciola hepatica infection had no “rim sign” or “target” sign, Liver abscesses were less than 3.0 cm in diameter, and the dilation of the bile duct were not observed. Conclusion Liver absessed caused by the fasciola hepatica infection have characteristic CT features. Combined with clinical examination and laboratory test, the reliability of diagnosis will be considerably increased.

Objective To express and purify the Schistosoma japonicum adenylate kinase(AK) protein of which the cDNA sequence was subcloned into the pET32a(+) soluble expression plasmid, and to evaluate its immunoreactivity. MethodsThe recombinant plasmid was transformed into {E.coli} BL21, and induced with IPTG for expression. The bacterial lysis was conducted by ultrasonication and the supernatant was analyzed by SDS_PAGE after boiling and centrifugation. The target protein was purified with the Ni_NTA agarose. The proteins on the gel were transferred to the PVDF film and then the immunoreactivity was tested by Western blotting using anti_6_His antibody, sera from rabbits 6 weeks after infected with cercariae or UV_attenuated cercariae. The purified protein was coated for ELISA to test the cercariae infected rabbit serum and the normal rabbit serum. Results The molecular weight of the target fusion protein was {Mr 40 000} after being induced with IPTG. The fused protein showed a single band when reacted with anti_6_His antibody, the cercariae infected rabbit serum and attenuated cercariae infected rabbit serum. Conclusion The AK protein is expressed as a fused protein with thioredoxin and its molecular weight is about {Mr 40 000}. This protein has a positive immunoreactivity with sera of rabbits infected with cercariae and UV_attenuated cercariae.

Objective To analyze the difference among antigens of Angiostrongylus cantonensis in different developmental stages and identify dominant diagnostic antigen for angiostrongyliasis. Methods Antigens of A.cantonensis in different developmental stages were analyzed by SDS-PAGE and immunoblot. Results The protein bands of all developmental stages were similar on SDS-PAGE. The Mr 40 000 , 50 000 , 66 000 and 80 000 antigens reacted not only with the sera of rats infected by A.cantonensis but also with the sera of normal rats. The Mr 104 000 antigen could be discerned by sera of rats infected with A.cantonensis for 2 weeks. The Mr 32 000 antigen could be recognized by sera of rats 2 weeks after infection, and the reaction became stronger with the infection continued. Conclusion The Mr 40 000 , 50 000 , 66 000 and 80 000 antigens might result in the unspecific reaction in the immunodiagnosis of angiostrongyliasis using the crude antigen of A.cantonensis. The Mr 104 000 of larva, Mr 33 000 of adult females and Mr 32 000 of the worms might be used as candidate antigens in early diagnosis and epidemiological survey of angiostrongyliasis.

AIM:To observe the expression of Toll like receptor 4 (TLR4) in the left ventricle of Goldblatt rats and to explore the role and mechanism of TLR4 in left ventricular remodeling of hypertension.METHODS:Goldblatt model of Two-kidney,one-clip (2K1 C) renovascular hypertension was induced in twenty-five rats(H group),and twenty rats served the sham-operated group(sham group).The tail cuff blood pressure was detected every week and echocardiogram was observed every other week.After eight weeks of operation,the rats were killed and the samples of the left ventricle were collected.The concentration of Ang Ⅱ in left ventricle was assessed by tadioimmunoassay.Western blotting and RT-PCR were used to exam the mRNA and protein expression of TLR4 in the left ventricle.Immunohistochemistry was adopted to exam the location of TLR4 in the myocardium.RESULTS:TLR4 mRNA and protein expression were consistently upregulated in the left ventricle of H group compared with sham group.In H group, predominantly sarcolemmal staining was observed,especially focal areas of intense TLR4 staining were found in juxtaposed regions of two or more adjacent myocytes;However,in sham group,TLR4 expression was diffuse and presumably cytoplasmic.Considerable correlation was found between blood pressure,MESS,LVMI,RWT,the concentration of Ang Ⅱ in left ventricle and the protein expression of TLR4 in myoeytes.CONCLUSION:During the development of left ventricular remodeling of Goldblatt rats,expression of TLR4 increases significantly.Enhanced expression of TLR4 locates in the sarcolemma,especially in juxtaposed regions of two or more adjacent myocytes.These indicate that TLR4 transmembrane receptor which is closely relative with inflammation and immunity probably contributes to the development of ventricular remodeling.[KEY WORDS]Hypertension;Ventricular remodeling;Receptors,Toll-like;Goldblatt rats

BACKGROUND: Antinuclear antibody, anticentromere antibody, anti-cytoplasm antibody, and antibody against SCL-70 are the main self-antibodies involved in systemic scleroderma (SSc) and are closely connected with the development of SSc.OBJECTIVE: To probe the value of various self-antibodies and proteins in evaluating the functional prognosis of patients with SSc. DESIGN: Case-control study.SETTING: Clinical Laboratory of Second Hospital Affiliated to Jiangxi Medical College.PARTICIPANTS: Totally 74 patients, 19 males and 55 females aged 12-59years old, were confirmed of SSc at the outpatient and inpatient departments of Second Hospital Affiliated to Jiangxi Medical College between December 1995 and December 2004. There were 46 cases of diffuse cutaneous SSc, 24 cases of localized cutaneous SSc, and 4 cases of overlapping syndrome. Meanwhile40 inpatients (14 males and 26 females aged 19-54years old) who received treatment due to other diseases were recruited from the same hospital.METHODS: The level of antinuclear antibody, anticentromere antibody,and anti-cytoplasm antibody was detected with indirect irnmuneofluorescence assay; antibody against SCL-70 was detected with Western blot; the level of serum immunoglobulin, C-reactive protein (CRP) and rheumatoid factor was examined with velocity dispersion turbidimetry.ticentromere antibody, anti-cytoplasm antibody, and antibody against SCL-70;RESULTS: Blood samples collected from the 74 patients with SSc and 40controls were proved eligible and all data entered the final statistical analanti-cytoplasm antibody, and antibody against SCL-70: It was obviously higher in SSc group than in control group [66% (49/74), 53% (39/74), 39%(29/74), 7% (5/74), 0, (x2=57.15, P ＜ 0.01)]. The positive rate of antinuclear antibody in patients with diffuse cutaneous SSc was significantly lower than in patients with localized cutaneous SSc [57% (26/46), 83%(20/24),(x2=5.03, P ＜ 0.05)], but the positive rate of antibody against SCL70 was significantly higher than localized cutaneous SSc [48% (22/46),rheumatoid factor in patients with SSc was markedly higher thanin control group [(16.89±11.94), (11.89±2.05) g/L; (23.06±6.18), (22.44±5.53) IU/mL,t=8.01, 2.46, P ＜ 0.01].CONCLUSION: The positive rate of antinuclear antibody, anticentromere antibody, anti-cytoplasm antibody, and antibody against SCL-70, as well as the level of immunoglobulin G and rheumatoid factor were obviously increased in patients with SSc, suggesting that these experimental parameters have the value in evaluating the prognosis of SSc.