Objective To prepare an attenuated Listeria vaccine Lmdd-LMP2A expressing the Ep-stein-Barr virus latent membrane protein 2A ( EBV-LMP2A) and evaluate its specific anti-tumor effects on nasopharyngeal carcinoma.Methods The gene fragment encoding EBV-LMP2A was amplified by PCR analysis and then subcloned into the shuttle vector p1565.PCR restriction enzyme digestion and DNA se-quencing were performed to identify the recombinant shuttle vector p1565-LMP2A.The p1565-LMP2A vector was then transformed into competent strains of the attenuated Listeria monocytogenes ( Lmdd) .The recombi-nant attenuated Listeria vaccine strain Lmdd-LMP2A was verified by Western blot assay.The histological sections of spleen and liver tissues were stained by Haematoxylin and eosin ( H&E) for analysis of inflamma-tion.A tumor-bearing HLA-A2 transgenic mouse model was established by subcutaneous injection of CNE-1/HLA-A2/LMP2A nasopharyngeal carcinoma cell line.The prepared Lmdd-LMP2A vaccine was inoculated into the mice via tail intravenous injection for the evaluation of specific CTL induction and the in vivo anti-tumor effects.Results The shuttle vector p1565-LMP2A and the recombinant attenuated Listeria vaccine strain Lmdd-LMP2A with stable expression of LMP2A protein were successfully constructed.The immunized mice showed mild inflammations with no structural damage and necrosis as indicated by H&E staining.The growth of tumors in tumor-bearing HLA-A2 transgenic mice was significantly inhibited by the immunization of Lmdd-LMP2A vaccine as compared with mice without inoculation.The survival time of mice was prolonged with the immunization of Lmdd-LMP2A vaccine.Conclusion The prepared attenuated Listeria vaccine Lm-dd-LMP2A showed specific anti-tumor effects with the safety advantage, suggesting the possibility of using it for anti-tumor therapy in clinic.

Objective To study the role of atrial natriuretic peptide (ANP) and renin-angiotensin-aldosterone system (RAAS) in the pathogenesis of salt-sensitive (SS) hypertension and mechanism of the hypotensive effect of benazepril and ANP in patients with SS essential hypertension. Methods Sixty-four patients with essential hypertension were divided into SS (n=30) and non-salt-sensitive (NSS, n=34) groups by modified Sulliran's method. Plasma ANP, angiotensin Ⅱ (AⅡ) and aldosterone (ALD) were determined before and during the period of salt loading test. Thirty healthy subjects as controls were also enrolled. A self-comparative study of benazepril with the placebo was performed in SS group. Before and after the placebo and benazepril therapy, blood pressure (BP) and plasma ANP were determined. Results (1)Basal plasma ANP level in the SS group was significantly lower than that in the NSS group. Basal plasma ANP level in the NSS group was also significantly lower than that in the control group [(110.28±15.40) pmol/L vs NSS (145.52±26.53) pmol/L and control (197.74±26.20) pmol/L]. Plasma ANP in both SS and NSS groups [(133.56±34.03) pmol/L and (169.20±35.91) pmol/L respectively, both P<0.05 vs control]. Percentage of increase of plasma ANP in SS and NSS groups was of no difference (P>0.05) . (2) No significant difference of basal plasma AⅡ and ALD levels were found between SS and NSS groups (P>0.05). There were no significant changes of plasma AⅡ and ALD during salt loading in both SS and NSS groups ( both P>0.05). (3) After the benazepril treatment, plasma ANP was increased significantly [(146.74±31.86) pmol/L , P<0.01]; both systolic and diastolic BP were reduced significantly in SS group. (4) Basal plasma ANP level was negatively correlated with the magnitude of increase of mean arterial pressure (MAP) by salt loading (b=-0.06, P<0.05). Conclusion Deficiency of circulating endogenous ANP may play an important role in the pathogenesis of SS hypertension. Benazepril could reduce BP and increase plasma ANP significantly in patients with SS hypertension.

APOBEC(apolipoprotein B mRNA-editing enzyme catalytic-polypeptide) family members were reported as innate immune molecules with anti-viral activity for many viruses, such as HIV and HBV.In order to understand the function of APOBEC, the APOBEC-3F and-3G were cloned, expressed, and the sub-cellular localization of them was detected.The genes of APBEC-3F and-3G were cloned from PHA-stimulated PMBC and expressed in the MDCK cell by transfection.The sub-cellular localization of APOBEC-3F and-3G were detected by immunofluorescence.APOBEC-3F and-3G were cloned by RT-PCR and confirmed by DNA sequencing.The immunofluorescence indicated APOBEC-3F and-3G were located in the cytosal.APOBEC-3F and-3G could inhibit HBV replication effectively in HepG2.2.15 cell.APOBEC-3F and-3G could not be trans-located into nuclear by nuclear location signal(NLS) or bi-NLS(B-NLS).These results will help the future research on the function of APOBEC.

Objective To discuss the effects of Radix Hedysari flavonoids on microvessel density (MVD) of pulmonary interstitial fibrosis model rats, and provide evidence for its development. Methods SPF Wistar rats were divided into blank group, model group, prednisone group, and Radix Hedysari flavonoids high-, medium- and low-dosage group. Pulmonary interstitial fibrosis model was established by intratracheal instillation of bleomycin. From the second day after model established, each drug treatment group was administered with corresponding drugs intragastrically at different points in time for 14 and 28 days. MVD was detected by immunohistochemistry. Results Neomicrovessel was increased significantly in pulmonary tissue of model rats of 14 days treatment, but slightly decreased in 28 days. MVD in 14, 28 days model group was significantly more than blank group (P<0.01). MVD in 14, 28 days Radix Hedysari flavonoids high dosage group was significantly less than model group (P<0.01). MVD in Radix Hedysari flavonoids medium-dosage group was between high-and low-dosage group, and MVD in medium-dosage group was similar with that in model group. Conclusion Radix Hedysari flavonoids could inhibit the formation of neomicrovessel in a dose dependent manner.

Objective:Increased reactive oxygen species (ROS) formation and by which in turn promotes cardiomyocytes apoptosis is associated with the pathogenesis and progression of various cardiac diseases. Small heat shock protein 27(Hsp27) could protect different cells from oxidative damage. By using tissue nonspecific overexpression Hsp 27 transgenic model, other investigators demonstrated that Hsp27 suppressed successfully kainate-induced seizures and hippocampal cell death in intact transgenic mice, and attenuated mimic ischemia/reperfusion injury in Langendorff-perfused isolated mice heart. As there are complicated and long distance neuro-humoral regulation associated with the development of cardiac diseases, it is better to choose a cardiac-specific overexpression transgenic model to study the effects of Hsp27 in hearts in vivo.Methods:A cDNA encoding human Hsp27 was subcloned into pBSII-SK+ containing the ?-myosin heavy chain (?-MHC) promoter (generously provided by Dr. J. Robbins, Children’s Hospital of Cincinnati, Ohio). BamHI-digested linear transgene consistent with the ?-MHC promoter, Hsp27 cDNA, and poly (A) of human growth hormone (hGH) was microinjected into the fertilized eggs from CBA/BL6 mice. Mice containing the transgene were identified by polymerase chain reaction. Founders revealed by this screening were used to establish independent transgenic lines. Following successful transmission, a range of tissues including heart, lung, liver, brain, skeleton muscle, spleen and kidney was screened by Western blot to confirm the cardiac specific expression of the transgene. Results and Discussion:Transgenic mice expressed Hsp27 under the control of a-MHC promoter. Cardiac tissues from independent TG line expressed abundant Hsp27, and as expected, Hsp27 expressed in cardiac tissues only, whereas none in liver, spleen, lung, kidney, brain and skeleton muscle. Surprisingly, Hsp25, the endogenous isoform of Hsp27 in murine, was downregulated by Hsp27 overexpression (data not shown), which is in contrast to the result of Hollander’s [1]. It needs to determine in future whether Hsp27 expression profile in heart could exert any effect on the regulation of Hsp25.Conclusion: The TG mice overexpression human Hsp27 specifically in the heart by using ?MHC- promotor were created. All TG mice expressed Hsp27 abundantly and cardiac-specifically. To our knowledge, it is the first report of creation of transgenic mice which overexpressing Hsp27 cardiac specifically. This current model suggests that the Hsp27 cardiac-specific over-expression of transgenic mouse remains a robust genetic tool for dissecting molecular and genetic events involving Hsp27, which could be a therapeutic target in heart failure.

BACKGROUND:Stroke is the leading cause of death and long-term disability. This study was undertaken to investigate the factors influencing daily activities of patients with cerebral infarction so as to take interventional measures earlier to improve their daily activities. METHODS:A total of 149 patients with first-episode cerebral infarction were recruited into this prospective study. They were admitted to the Encephalopathy Center, Department of Neurology, the First Affiliated Hospital of Wenzhou Medical College in Zhejiang Province from August 2008 to December 2008. The baseline characteristics of the patients and cerebral infarction risk factors on the first day of admission were recorded. White blood cell (WBC) count, plasma glucose (PG), and many others of laboratory targets were col ected in the next morning. Barthel index (BI) was calculated at 2 weeks and 3 months respectively after onset of the disease at the outpatient clinic or by telephone cal . Lung infection, urinary tract infection and atrial fibrillation if any were recorded on admission. The National Institute of Health Stroke Scale (NIHSS) scores and the GCS scores were recorded within 24 hours on and after admission, at the second week, and at the third month after the onset of cerebral infarction respectively. RESULTS:The factors of BI at 2 weeks and 3 months after onset were the initial PG level, WBC count and initial NIHSS scores. Besides, urinary tract infection on admission was also the factor for BI at 3 months. CONCLUSION:Active measures should be taken to control these factors to improve the daily activities of patients with cerebral infarction.

Objective To explore the effect of electroacupuncture combined with rehabilitation on P300 auditory event-related potentialsin patients with cerebral infarction. Methods 105 patients with cerebral infarction were divided into 3 groups with 35 cases in each group.The comprehensive rehabilitation group received routine rehabilitation and electroacupuncture, the general rehabilitation group only receivedroutine rehabilitation, and the control group received no intervention. They were treated once a day, 10 times as a course. P300 wasdetected before and 3 courses after treatment. Results The latency was shorter and the amplitude was higher in the comprehensive groupthan in the general group (P<0.05). Conclusion Electroacupuncture combined with rehabilitation technology can improve cognitive functionof patients with cerebral infarction.

Objective To explore the effect of electroacupuncture combined with rehabilitation technology on the recovery of motor function in patients with cerebral infarction. Methods 105 patients with cerebral infarction were divided into 3 groups with 35 cases in each group. The comprehensive rehabilitation group received routine rehabilitation and electroacupuncture therapy, the general rehabilitation group only received routine rehabilitation, the control group received no intervention. They were treated once a day, 10 times as a course.Fugl-Meyer Assessment (FMA) was used to assess the motor function after 3 courses. Results The scores of FMA were higher in both rehabilitation groups than in the control group (P<0.05), and it was higher in the comprehensive group than in the general group (P<0.05). Conclusion Electroacupuncture combined with modern rehabilitation technology could facilitate to recover motor function in patients with cerebral infarction.

OBJECTIVETo find the objective indices for traditional Chinese medicine (TCM) pattern identification in women patients with climacteric hypertension (CH) by exploring the relationship between TCM pattern with blood levels of sex hormone, lipid and glucose.

METHODSTCM syndrome differentiation was made in 75 women CH patients, 43 as yin-deficiency with inner dryness pattern (YDD) and 32 as non-YDD, and correlation analyses were conducted on TCM patterns and blood levels of estradiol (E2), lipids and glucose.

RESULTSBesides the level of E2 in YDD was lower than that in non-YDD [(112.26 -/+ 66.81) pmol/L vs (146.69 +/- 83.59) pmol/L, P < 0.05), no difference between the two patterns was found in the other indices measured (P > 0.05). Further subunit analysis showed that in patients with CH grade 1, blood levels of total cholesterol (TC), high density lipoprotein (HDL) and apoprotein A-I (ApoA-I) were higher in the 13 patients of YDD than those in the 9 of non-TDD (P < 0.05); in patients with CH grade 2, levels of E2, progesterone (T), HDL and ApoA-I were lower but level of follicle stimulating hormone (FSH) was higher in the 20 YDD than those in the 10 non-YDD (all P < 0.05); while in patients with CH grade, no statistically significant difference was shown between 3 patients of 10 with YDD and 13 with non-YDD in all the indices measured (P > 0.05).

CONCLUSIONBlood level of E2 shows different characteristics between CH patients of YDD and non-YDD patterns, suggesting that it may provide some help to syndrome micro-differentiation, and taking it as an objective index for syndrome typing of climacteric hypertension in female is considerable.

Adult ; Blood Glucose ; metabolism ; Estradiol ; blood ; Female ; Humans ; Hypertension ; blood ; Lipids ; blood ; Menopause ; blood ; Middle Aged ; Progesterone ; blood ; Yin Deficiency ; blood

Objective To investigate the relationship between the promoter of IL-12B gene polymorphism and the susceptibility and clinical features of chronic gastritis and duodenal ulcer with or without Helicobacter pylori(Hp) infection in children and adolescent.Methods Mucosal biopsies were obtained from 132 children and adolescent (patient group),including 100 children with chronic gastritis and 32 children with duodenal ulcer,undergoing an upper gastrointestinal endoscopy for dyspeptic symptoms.Biopsy specimens were stained with hematoxilin and eosin (HE),and gastritis was graded according to the Sydney system.Serology,urease test and histology were taken to assess Hp status.Genomic DNA was obtained from peripheral blood or gastric biopsies of patients and 102 healthy children as normal control group.The promoter of IL-12B +1188A/G gene polymorphism was genotyped by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and sequencing.The genotype distributions and allele frequencies were compared between the study group and the normal control group,and the association of genotypes with clinicopathological features was studied.IL-12B mRNA level expressions in gastric mucosa were confirmed by reverse transcription PCR biopsy-based tests.Results The genotype distributions and allele frequencies of IL-12B +1188A/G gene polymorphisms were similar in gastric upper gastrointestinal diseases and healthy subjects.The IL-12B +1188A/G gene polymorphisms were not associated with Hp status.IL-12B+1188A/G gene polymorphisms did not affect IL-12B mRNA level expressions and were not associated with the degree of antrum chronic inflammation.Conclusions These data suggest that IL-12B+1188A/G gene polymorphisms are not associated with susceptibility to chronic gastritis and duodenal ulcer in children and adolescent.