Objective To Clone,express,purify 38 ku protein of mycobacterium tuberculosis,to study its immunological characteristics,and to evaluate its potenial value for serodiagnosis of tuberculosis.Methods Extract DNA from standard strain of H37Rv as the template,amplify gene of 38 ku protein by PCR,insert to PET-30a(+)and construct the recombinant plasmid,express 38 ku protein in E.coli BL21(DE3),purify by Nickel affinity chromatography,at last get target proteins of higher purity,analyze its immol/Lunological characteristics by Western blotting and ELISA technology from Feb.2003 to Mar.2004. Results The clone was analyzed at the nucleotide lever and showed the same DNA sequence coding for natural 38 ku protein. The recombinant protein expressed in inclusion body in E.coli BL21(DE3). The purity of terget protein was 92.7% by Nickel affinity chromatography,Western blotting assays showed that the recombinant protein had satisfactory antigenicity . 38 ku protein detected TB postive and negative refference serum based on the mechanism of indirect ELISA,results showed that the sensitivity was 80.5%(33/41)and the specificity reached to 96%(25/26).Conclusion The recombinant protein expressed in inclusion body in E.coli BL21(DE3)and had satisfactory antigenicity,and might be selected as one of serodiagnostic antigen of tuberculosis.

Based on the conservation of nucleotides at splice sites, the characteristics of base composition and base correlation in the adjacent segment sequences, the distance between alternative donor or acceptor splice sites and the content of GC and TC near splice sites, the donor and acceptor splice sites for alternative and constitutive introns are predicted by use of the method of diversity measure combined with quadratic discriminant analysis. For alternative splice sites the total prediction accuracies are 87.9% and 89.9% for donors and acceptors respectively (with the chosen threshold -2). For constitutive splice sites the total accuracy are 92.8% and 94.3% for donors and acceptors respectively (with the chosen threshold -1).

Based on the characteristic of nucleotide distribution in nucleosome positioning and inhibiting sequences, the method of Increment of Diversity with Quadratic Discriminant (IDQD) was applied to the classification of these two types of sequences. The mean area under ROC curve archives 0.958. By using this model, the nucleosome formation potential was analyzed in the regions around the splice sites (GT/AG). The results show that coding regions have a high potential to form the nucleosome and the primary RNA transcripts are rigid, while DNA sequences corresponding to the splice sites and their adjacent intron regions tend to be nucleosome free and the primary transcripts from these regions are relative flexible. Moreover, the negative correlation between nucleosome positioning/inhibiting of DNA sequences and RNA flexibility/rigidity is demonstrated around the splice sites, providing a mechanism for understanding the correlation between the nucleosome positioning of DNA and the splicing of transcribed RNA sequences.

OBJECTIVE: To investigate the expression of EBV-encoded latent membrane protein 2A (LMP2A) and epithelial-mesenchymal transformation(EMT) associated markers (E-cadherin and fibronectin) in nasopharyngeal carcinoma (NPC) and its clinical significance. METHOD: The expression of LMP2A, E-cad-herin and fibronectin proteins in 32 cases of chronic nasopharyngeal inflammation, 56 cases of NPC and 18 cases of NPC lymph node metastasis were examined byimmunohistochemical SP method. RESULT: (1)The positive rates of LMP2A in NPC and its lymph node metastasis were significantly higher than those of chronic nasopharyngeal inflammation (89. 3%vs 37. 5%o and 77. 8% vs 37. 5%) respectively (both P<0. 01); The normal expression rates of E-cadherin in NPC and its lymph node metastasis were significantly lower than those of chronic nasopharyngeal inflammation (33. 9% vs 90. 6% and 5. 6% vs 90. 6%) respectively (both P<0. 01); The positive rates of fibronectin in NPC and its lymph node metastasis were significantly higher than those of chronic nasopharyngeal inflammation (83. 9% vs 28. 1% and 72. 2% vs 28. 1%) respectively (both P<0. 01). (2) ZLMP2A expression were negatively correlated with normal expression of E-cadherin (r= -0. 387, P<0. 01), and were positively correlated with fibronectin (r= 0. 421, P<0. 01). (3)LMP2A, E-cadherin and fibronectin expression were significantly correlated with N stage and clinical stage (both P<0. 05), but the three proteins were not significantly correlated with M stage (both P> 0. 05). In addition, LMP2A and E-cadherin expression were significantly correlated with T stage (both P<0. 01). CONCLUSION LMP2A and fibronectin expressions were increased in NPC, but normal expression of E-cadherin were decreased. LMP2A may promote lymph node metastasis and malignant progression of NPC by induce EMT through downregulation of E-cadherin and upregulation of fibronectin.
Cadherins ; biosynthesis ; Carcinoma ; Epithelial-Mesenchymal Transition ; Fibronectins ; biosynthesis ; Humans ; Lymphatic Metastasis ; Membrane Proteins ; biosynthesis ; Nasopharyngeal Carcinoma ; Nasopharyngeal Neoplasms ; metabolism ; pathology ; Nasopharynx

Objective To explore the effect of chronic intermittent hypoxia (CIH) on hippocampus neuron and growth associated protein-43 (GAP-43) in rats. Methods 40 adult male Sprague-Dawley rats were randomly divided into control group, CIH 1 week group, CIH 3 weeks group and CIH 5 weeks group with 10 rats in each group. The CIH groups were exposed to intermittent hypoxia in designed cabin 8 hevery day for 1 week, 3 weeks and 5 weeks. The pathological changes of hippocampal neurons were observed by HE staining. The GAP-43 expression of hippocampus was detected by immunohistochemical staining. Results There was degeneration and necrosis of hippocampal neurons in the CIH rats, and it became worse gradually along with the hypoxia. Compared with the control group, the GAP-43 expression in hippocampus increased in CIH rats. The peak of GAP-43 expression appeared in the 1st week (P=0.038) and then it decreased in the 3rd week (P=0.382) and it was closed to the normal level in the 5th week (P=0.860). Conclusion Degeneration and necrosis of hippocampal neurons exacerbated gradually along with the hypoxia, while GAP-43 expression changed inconsistently.

Objective To establish the chromatographic fingerprint of mushroom polysaccharides by 1-phenyl-3-methyl-5-pyrazolone( PMP) pre-column derivatization. Methods The mushroom polysaccharides was extracted by hot distilled water, precipitated by alcohol, and hydrolyzed into monosaccharides by Trifluoroacetic Acid (TFA). The hydrolysate was derivatized with 1-phenyl-3-methyl-5-pyrazolone ( PMP ) and tested via HPLC to study the monosaccharide components in mushroom polysaccharides. Results Fingerprint was established with 13 common peaks, 6 peaks in which were identified as mannose, D-glucuronic acid, D-glucose, galactose, xylose and L-fucose. The glucose accounted for the most, followed by mannose, galactose and fucose. Conclusion Development of fingerprint chromatogram by HPLC is a stable, simple, and repeatable way, which can be applied to the quality control of mushroom polysaccharides.

Objective To evaluate the effect of electro-acupuncture (EA) at Zusanli and Feishu on endotoxin shock-induced acute lung injury in rabbits.Methods Sixty healthy male New Zealand white rabbits aged 2 months weighing 1.5-2.0 kg were randomly divided into 6 groups (n =10 each):group sham operation (group S); group zinc protoporphyrin-Ⅸ (ZnPP-Ⅸ) (group Z); group lipopolysaccharide (LPS) (group L); group LPS + EA (group EL) ; group LPS + sham EA (group SEL) and group LPS + EA + ZnPP-Ⅸ (group ELZ).The animals were anesthetized with intraperitoneal 10％ chloral hydrate 400 mg/kg and tracheostomized.The animals kept spontaneous breathing.Right internal carotid artery was cannulated for BP monitoring.Ear vein was cannulated for drug administration.LPS 5 mg/kg was injected iv in groups L,EL,SEL,ELZ.Endotoxin shock was confirmed by decrease in BP by 20 ％ of the baseline value and PaO2/FiO2 ≤ 300.ZnPP-Ⅸ (heme oxygenase (HO-1 ) inhibitor)10μmol/kg was injected intraperitoneal at 2 h after LPS injection in groups Z and ELZ.Bilateral 15 min EA stimulation of Zusanli and Feishu ( according to atlas of animal acu-points) was performed once a day for 5 days before LPS administration in groups EL and ELZ.The animals were sacrificed by blood-letting at 6 h after LPS administration.The lungs were removed for microscopic examination (0 =no injury,4 =most severe injury),detection of alveolar epithelial cell apoptosis (by TUNEL) and determination of HO-1 protein and mRNA expression.Results LPS significantly increased lung injury scores,alveolar epithelial cell apoptosis index (the number of apoptotic cells/total cells) and HO-1 protein and mRNA expression.EA significantly attenuated lung injury and alveolar epithelial cell apoptosis induced by LPS and further increased the expression of HO-1 protein and mRNA in group EL as compared with group L.The protective effects of EA was counteracted by ZnPP- Ⅸ in group ELZ.Conclusion EA at Zusanli and Feishu can attenuate endotoxin shock-induced lung injury by up-regulation of HO-1 expression and inhibiting alveolar epithelial cell apoptosis in the lung.

Objective To evaluate comprehensively the effect of vibratory sputum ejectionmachine on patients by comparing the volume of vibratory sputum ejection machine and the traditional back percussion sputum expectoration.Methods Searching thoroughly Chinese Journal Full-text Database(CJFD),VIP CJFD,Wanfang Periodical Database,collecting the clinical randomized controlled trials of domestic vibratory sputum ejection machine and traditional back percussion sputum expectoration and employing RevMan5.0 processing software for statistical analysis.Results Vibration expectoration instrument is better than the traditional back percussion sputum expectoration.Conclusions Vibratory sputum ejection machine can increase the amount of expectoration drainage in patients and reduce pulmonary infection.

Objective To understand the radiation protection effect of pre-irradiation administrations of nilestriol on the mice with bone marrow type of acute radiation syndrome after irradiation with 60Co γ-rays,along with its mechanisms for improvement of hematopoiesis.Methods The nilestriol administration protocols were prepared by analysis of peripheral blood cell counts and survival rate experiment on mice.The mechanisms by which the pre-irradiation twice administrations improved the post-irradiation recovery of bone marrow hematopoiesis were studied by the analysis of the surface marker of bone marrow hematopoietic stem/progenitor cells of mice and by the inspection of hematopoietic progenitor cell colony and by using histopathological assessment of bone marrow.Results Pre-irradiation administration of nilestriol at two-or three-day intervals had been shown to increase survival rates up to 100％ in mice exposed to 9.0 Gy γ-rays,which was superior to a single administration (20％,x2 =21.66,21.66,P ＜0.05).The pre-irradiation administration both at one-day or two-day intervals were capable of improving the recovery of peripheral blood counts,including white blood cell (WBC),red blood cell (RBC),and platelet in mice exposed to 6.5 Gy (F =21.33,100.9,49.34,19.19,P ＜ 0.05),showing the better effects than a single administration (F =17.11,63.38,21.89,14.37,P ＜ 0.05).The two-day-interval administration of nilestriol could significantly increase the numbers of bone marrow hematopoietic stem/progenitor cell counts (t =8.58,2.80,P ＜ 0.05) in mice on day 10 after 6.5 Gy irradiation.This also could be capable to significantly improve colony formation,with there being statistical difference compared with single administration(t =4.29,6.34,P ＜ 0.05).Also the administration at two-day-interval were also usefull in reconstruction of hematopoietic cell hyperplasia of bone marrow of irradiated mice.Conclusions As compared with conventional single admination,the pre-irradiation multiple administrations of nilestriol showed significantly improved radiation protection effects.Considering a nuclear medical emergency rescue,it is recommended to follow the pre-irradiation administration of nilestriol at two-day interval,which could obtain the best protection effects at minimum administration frequency.

Objective To evaluate the efficacy and safety of biphasic insulin aspart 30 (BIAsp30)plus metformin in type 2 diabetes subjects switching from basal insulin plus oral antidiabetic drugs (OAD)Methods During 16 weeks, multiple-center, open-label, and single-arm study including 2 weeks of screening period,4 weeks of run-in period,and 16 weeks of treatment period were carried out. Subjects with type 2 diabetes mellitus inadequately controlled on basal insulin therapy with or without oral antidiabetic drugs were switched to twice-daily BIAsp30 plus metformin with dose titration to achieve fasting plasma glucose target. Results Of the 293 Chinese subjects exposed to trial drugs [age: ( 54.0±9.6 ) years, diabetes duration: ( 8.54±5.49 ) years, body mass index: (24.89±3.28)kg/m2, baseline HbA1c: 8.16% ±0.89%], 122 were previously treated with basal insulin analogues and 169 with human basal insulin. At end of the trial ,the mean reduction of HbA1 c was 1.30% ±0.96% (P＜0. 01 ). The proportion of patients achieved HbA1c＜7.0% and HbA1c ≤6.5% was 60.4% and 38.9% respectively. 8-point plasma glucose measurements showed significant improvements at all the time points examined ( all P＜0. 01 ) ,and the average value of all 8 points measured decreased from ( 10.53±2.58 ) mmol/L atbaseline to (7.79± 1.58 ) mmol/L at the end of treatment ( P＜0. 01 ), reduced by 2.76 mmol/L. Postprandial glucose increments were significantly reduced after breakfast ( -1.73 mmol/L,P＜0.01 )and dinner ( -1.28 mmol/L,P＜0.01 ), while no significant reduction was observed after lunch ( -0.09 mmol/L, P = 0. 734 5 ). No severe adverse effect and no major hypoglycemia were reported. The overall hypoglycaemia rate was 2.68 events/subject year. The average weight gain was (0. 76 ±0. 14 )kg (P＜0. 0l ). Conclusion Twice-daily BIAsp30 plus metformin is effective and safe to type 2 diabetic subjects inadequately controlled on basal insulin treatment.BIAsp30 treatment should be considered for type 2 diabetic subjects who have unsatisfactory response to previous basal insulin treatment.