Objective] To explore the humoral and cellular immune responses in mice to eukaryotic expression recombinant plasmid encoding histidine rich protein 2 (HRP\|Ⅱ) of Plasmodium falciparum. [Methods] The start and stop codes were introduced into HRP\|Ⅱ gene fragment, the reading frame and the position of start and stop codes in HRP\|Ⅱ were identified by sequencing. HRP\|Ⅱ fragment containing the start and stop codes was cloned into pcDNA3 1(\|) to form pcDNA3 1(\|)/HRP\|Ⅱ. The BALB/c mice were immunized i.m. with the plasmids for 3 times in 3 weeks intervals. Two weeks after the last immunization, the sera and splenocytes were collected to investigate anti\|HRP\|Ⅱ antibodies by ELISA and the splenocytes proliferation response to HRP\|Ⅱ. [Results] Sequence data show that the reading frame and the position of start and stop codes are correct. Restriction enzyme digestion indicated that the HRP\|Ⅱ gene fragment containing start and stop codes was successfully cloned into pcDNA3 1(\|). Mice raised significant anti\|HRP\|Ⅱ antibodies after pcDNA3 1(\|)/HRP\|Ⅱ immunization, and the splenocytes proliferated prominently when stimulated with HRP\|Ⅱ protein. [Conclusion] Eukaryotic expression recombinant plasmid \{encoding\} HRP\|Ⅱ gene can induce significantly humoral and cellular immune response in mice. HRP\|Ⅱ gene may be a good candidate for P.falciparum blood\|stage multiple DNA vaccine.

OBJECTIVEEstablish the model of mouse with chronic hepatitis B virus (HBV) and nonalcoholic fatty liver disease (NAFLD).

METHODSTake 100 HBV transgenic, BALB/c mice of 4 weeks old, with each gender half. Then pick out 70 mice in one group to feed high-fat feed and the rest to feed normal feed. At the end of week 16, random kill 10 mice of high-fat, then liver tissue and serological detection target identification model is established in this paper. After that, divide the mice into model group and comparison group with 30 mice in each group. Feed model group with high-fat feed, comparison group with normal feed and normal group with normal feed till week 72 (including previous 16 weeks). Kill 10 mice of each group at the end of week 24, 48 and 72 respectively, fully automatic biochemical instrument detection of serum ALT, AST, TC, TG, FBG, fluorescence quantitative PCR method to detect HBV-DNA, chemiluminescence detection of HBsAg, liver biopsy after HE staining to evaluate histology change, observe mice model of dynamic evolution.

RESULTS(1) Feed high fat feed after 16 weeks, mice's weight, serum ALT, AST, TC, TG, FBG and blood biochemical indicators increased, HBV-DNA positive, liver HE staining obviously big blister fatty degeneration of liver cells and within the lobule lymphocytes infiltration, NAFLD activity score (NAS) getting close to NASH, the model of chronic HBV carries with NAFLD mouse built successfully. (2) The TC and TG values of model group in each period were higher than that of comparison group and normal group. (3) In week 24 and 72, HBV-DNA values of each group are obvious different from the other two groups and the difference can be applied to statistical significance (P < 0.05). (4) In week 48 and 72, NAS of each group are obvious different from the other two groups and the difference can be applied to statistical significance (P < 0.05).

CONCLUSIONS(1) Chronic HBV carries with NAFLD mice model can be established by HBV transgenic mice fed by high fat feed. (2) NAFLD accelerates the liver disease of the mice carrying HBV to some extent.

Animals ; Disease Models, Animal ; Fatty Liver ; complications ; pathology ; virology ; Female ; Hepatitis B virus ; genetics ; isolation & purification ; physiology ; Hepatitis B, Chronic ; complications ; pathology ; virology ; Humans ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Transgenic ; Non-alcoholic Fatty Liver Disease

OBJECTIVETo determine the prevalence of dental anxiety (DA) in patients with cardiovascular diseases before tooth extraction so as to provide psychotherapy.

METHODS144 patients with cardiovascular diseases were asked to complete a questionnaire modified from Corah Dental Anxiety Scale (DAS) while awaiting for tooth extraction.

RESULTSThe incidence rate of DA in these patients was 7.6%. Anxiety level on anesthesia procedure was the highest. DAS score was higher among patients with the following factors: inactive perception, female, below 60 years old, experiences of painful tooth extraction, a documented history of single cardiovascular disease, under ECG monitoring for the first time and complicated tooth extraction.

CONCLUSIONDA existed in patients with cardiovascular diseases before tooth extraction and was related to many factors. Medical staff should take necessary measures to provide them with psychotherapy and nursing guidance before tooth extraction.

Adult ; Aged ; Aged, 80 and over ; Cardiovascular Diseases ; complications ; psychology ; China ; epidemiology ; Dental Anxiety ; epidemiology ; Female ; Humans ; Male ; Middle Aged ; Prevalence ; Surveys and Questionnaires ; Tooth Diseases ; complications ; surgery ; Tooth Extraction ; psychology

OBJECTIVETo investigate the beneficial effects of Rhein (RH) on hepatic progression in hepatitis B virus (HBV)-transgenic mice with nonalcoholic steatohepatitis induced by a high-fat (HF) diet.

METHODSA mice model of HBV chronic infection concomitant with liver steatosis was induced by a HF diet in 4-week old HBV-transgenic mice for 16 weeks (n = 130). Thereafter, the mice were divided randomly into control group (back to normal chow), model group (continuing HF diet), RH group [continuing HF diet and administering with 120 mg/(kg x d) RH by gavage] and Essentiale group [continuing HF diet and administering with 69.2 mg/(kg x d) Essentiale by gavage] with 30 mice in each, and were sacrificed at the end of 24-week and 48-week respectively. Serum levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), total cholesterol (TC), triglyceride (TG) and fasting plasma glucose (FPG) were measured by an automatic biochemical analyzer, and serum HBV-DNA was determined with qPCR. Hepatic histology was evaluated by HE staining with a light microscope.

RESULTS(1) An histological change composed of steatosis, lymphocytes intralobular infiltration and ballooning was observed after 48 weeks feeding of HF diet, in part mimicking that of NASH patients as evidenced by a NAFLD activity score (NAS) of 3.58 +/- 1.44 points. (2) Histologically, the NAS of model group was higher than that of control group at both time points. RH failed to lessen NAS whereas Essentiale improved the NAS at 48-week. (3) Serum levels of TC, TG and FPG were significantly different between 4 groups at 24-week, with a comparable low value in both RH and Essentiale group. A similar change was evident at 48-week. (4) In terms of HBV viral load, a significantly lower level in Essentiale group than the others was observed at both time points.

CONCLUSIONHF diet feeding is able to induce a mouse model of HBV chronic infection concomitant with NASH. RH is effective in alleviating the glucose and lipid metabolism but ineffective in improving the hepatic histology in this model, in contrast, backing to normal chow achieved a better effect in this aspect.

Animals ; Anthraquinones ; administration & dosage ; Diet, High-Fat ; adverse effects ; Disease Models, Animal ; Disease Progression ; Fatty Liver ; complications ; etiology ; metabolism ; prevention & control ; Female ; Glucose ; metabolism ; Hepatitis B virus ; physiology ; Hepatitis B, Chronic ; complications ; metabolism ; virology ; Humans ; Lipid Metabolism ; drug effects ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Transgenic ; Non-alcoholic Fatty Liver Disease

OBJECTIVETo investigate the expression of F4/80, NF-kappaB, p-AKT, AKT in the liver of nonalcoholic fatty liver disease (NAFLD) mice. To determine the role of Kupffer cells (KCs) in the development of NASH (non-alcoholic steatohepatitis), and understand the pathogenic mechanism of NASH.

METHODSFive C3H/HeN mice fed with normal diet were served as controls, while fifteen fed with high fat, high fructose, high fat combined fructose diet respectively for 16 weeks were as NAFLD mice models. The liver inflammation and hepatic damage were examined, and the expression of F4/80, NF-Kb, p-AKT, AKT and the content of lipid in the liver were also detected.

RESULTSChronic intake of high fat and 30% fructose solution caused a significant increase in hepatic steatosis in animals in comparison to water controls. Liver F4/80 and NF-kappaB were significantly higher in high fat and high fat combined fructose diet fed mice than that in controls (P < 0.01, P < 0.01), F4/80 protein were higher in high fat diet treated mice than those in fructose and high fat combined fructose groups (P < 0.01, P < 0.01). Markers of insulin resistance (e. g, hepatic phospho-AKT, AKT) were only altered in fructose-fed or high fat combined fructose animals (P < 0.01, P < 0.01).

CONCLUSIONHigh fat and fructose diet may induce NAFLD in C3H/HeN mice. Kupffer cells and signal pathway proteins were activated, and they may play key roles in the initiation and progression of NASH.

Animals ; Diet, High-Fat ; adverse effects ; Fatty Liver ; etiology ; immunology ; metabolism ; Female ; Fructose ; adverse effects ; Humans ; Kupffer Cells ; immunology ; Lipid Metabolism ; Liver ; immunology ; metabolism ; Male ; Mice ; Mice, Inbred C3H ; NF-kappa B ; immunology ; Non-alcoholic Fatty Liver Disease ; Oncogene Protein v-akt ; immunology ; Signal Transduction

OBJECTIVETo investigate the effect of RNA interfering TLR4 signal pathway on phagocytosis of Kupffer cells.

METHODSRAW2647 mice mononuclear macrophage leukemia cells were observed. The tested group was interfered by Tlr4-mus-1567 RNA which had the best result confirmed by QPCR, cells interfered by Negative Control RNA as NC group, and normal cell as control. We perform the phagocytosis test on each group.

RESULTSThe tested group has lower phagocytes percentage than control (17.67% +/- 3.51% vs 32.00% +/- 3.00%, P < 0.01), and lower phagocytic index (46.33% +/- 7.51% vs 82.00% +/- 6.08%, P < 0.01).

CONCLUSIONSDecreased phagocytic activity was observed on Kupffer cells by RNA interference.

Animals ; Kupffer Cells ; immunology ; Mice ; Phagocytosis ; RNA Interference ; Signal Transduction ; Toll-Like Receptor 4 ; genetics ; immunology

OBJECTIVETo evaluate efficacy of closed reduction and percutaneous hollow screw fixation with prototypical retractor in treating calcaneal fracture.

METHODSFrom January 2009 to June 2011, 39 patients (43 feet) with calcaneal fracture were treated by closed reduction and percutaneous hollow screw fixation with prototypical retractor. There were 33 males and 6 females, aged from 19 to 61 years with an average of 36.7 years. According to type of Sanders, type II were in 19 feet, type III were in 24 feet. Preoperative and postoperative X-ray were estimated, the data of height, width, Bölher angle, Gissane angle of calcaneous were collected, and ankle function were estimated according to AOFAS system.

RESULTSAll patients were followed up from 6 to 36 months with an average of (15.4 +/- 3.1) months. All wounds were healed well, no skin edge necrosis and infections occurred. Before operation, the height of calcaneous was average of (32.45 +/- 3.51) mm, width was (41.60 +/- 2.42) mm, Bölher angle was (8.64 +/- 13.2) degrees and Gissan angle was (136.35 +/- 15.23) degrees; while after operation, the height of calcaneous was average of (43.62 +/- 1.02) mm, width was (38.02 +/- 1.28) mm, Bölher angle was (26.87 +/- 5.32) degrees and Gissan angle was (120.78 +/- 5.34) degrees, and had significanty differences between preoperative and postoperative treatment (P<0.05). AOFAS score was improved from preoperative (35.64 +/- 11.23) to postoperative (76.18 +/- 9.87), and 29 cases got excellent results, 11 good and 3 fair. COMCLUSION: Closed reduction and percutaneous hollow screw fixation auxiliary by the retractor for the treatment is a good way, which has advantages of simple operation, satisfactory reduction fixation, reliable fixation, minimally invasive, less complications and rapid recovery.

Adult ; Bone Screws ; Calcaneus ; diagnostic imaging ; injuries ; surgery ; Female ; Follow-Up Studies ; Fracture Fixation, Internal ; instrumentation ; Fractures, Bone ; diagnostic imaging ; surgery ; Humans ; Male ; Middle Aged ; Skin ; Tomography, X-Ray Computed ; Treatment Outcome ; Young Adult

BACKGROUNDA growing number of studies worldwide have advocated the replacement of whole-breast irradiation with accelerated partial breast irradiation using three-dimensional conformal external-beam radiation (APBI-3DCRT) for early-stage breast cancer. But APBI can be only used in selected population of patients with early-staged breast cancer. It is not replacing the whole breast radiotherapy. This study aimed to examine the feasibility and acute normal tissue toxicity of the APBI-3DCRT technique in Chinese female patients who generally have smaller breasts compared to their Western counterparts.

METHODSFrom May 2006 to December 2009, a total of 48 Chinese female patients (with early-stage breast cancer who met the inclusion criteria) received APBI-3DCRT after breast-conserving surgery at Sun Yat-sen University Cancer Center. The total dosage from APBI-3DCRT was 34 Gy, delivered in 3.4 Gy per fractions, twice per day at intervals of at least six hours. The radiation dose, volume of the target area and volume of irradiated normal tissues were calculated. Acute toxicity was evaluated according to the Common Toxicity Criteria (CTC) 3.0.

RESULTSAmong the 48 patients, the planning target volume for evaluation (PTVE) was (90.42 ± 9.26) cm³, the ipsilateral breast volume (IBV) was (421.74 ± 28.53) cm³, and the ratio between the two was (20.74 ± 5.86)%. Evaluation of the dosimetric characteristics of the PTVE revealed excellent dosimetric results in 14 patients and acceptable results in 34 patients. The dose delivered to the PTVE ranged from 93% to 110% of the prescribed dose. The average ratio of the volume of PTVE receiving 95% of the prescription dose (V95) was (99.26 ± 0.37)%. The habituation index (HI) and the conformity index (CI) were 1.08 ± 0.01 and 0.72 ± 0.02, respectively, suggesting good homogeneity and conformity of the dose delivered to the target field. The radiation dose to normal tissues and organs was within the dose limitation. Subjects experienced mild acute toxicity. The main manifestations were breast edema in 22 patients, breast pain in 7, skin erythema in 25, general malaise in 22 and cytopenia in 8. No acute radiological cardiac or pulmonary injury was found.

CONCLUSIONSThe results of our short-term follow-up showed that it is feasible to perform APBI-3DCRT for early-stage breast cancer after breast-conserving surgery in Chinese female patients with smaller breasts. However, further studies are required to elucidate its efficacy and long-term side effects.

Adult ; Asian Continental Ancestry Group ; Breast Neoplasms ; radiotherapy ; surgery ; Female ; Humans ; Middle Aged ; Radiotherapy, Conformal ; adverse effects

OBJECTIVETo study the radiologic and pathologic features of primary intermediate hemangioendothelioma of the bone.

METHODSFive cases of primary intermediate hemangioendothelioma of bone encountered in the past three years were enrolled into the study. The clinical, radiologic, pathologic and immunohistochemical features of the tumors were reviewed.

RESULTSThe patients included 3 children with Kaposiform hemangioendothelioma and 2 elderly with retiform hemangioendothelioma. Four of the cases affected long bones and the remaining case affected the clavicle. One case showed multifocal involvement of the humerus. Radiologically, the tumors showed borderline to low-grade bony destruction, with various degrees of cortical defect. Intralesional or perilesional bone formation was demonstrated in 4 cases and radial spicules were seen in 1 case. The histopathologic features of primary intermediate hemangioendothelioma of bone were similar to those of soft tissue, except for the presence of reactive bone formation. Immunohistochemically, the tumor cells were positive for CD31 (5/5), CD34 (5/5), vimentin (5/5) and smooth muscle actin (3/5) but negative for cytokeratin and epithelial membrane antigen.

CONCLUSIONSPrimary intermediate hemangioendothelioma of bone is a distinct entity and similar histologic classification applies as in its soft tissue counterparts. Comparison of the biologic behavior requires long-term follow-up studies.

Actins ; metabolism ; Antigens, CD34 ; metabolism ; Bone Neoplasms ; diagnostic imaging ; metabolism ; pathology ; Child ; Clavicle ; pathology ; Diagnosis, Differential ; Female ; Femur ; pathology ; Hemangioendothelioma ; diagnostic imaging ; metabolism ; pathology ; Hemangiosarcoma ; pathology ; Humans ; Humerus ; pathology ; Immunohistochemistry ; Infant ; Kasabach-Merritt Syndrome ; diagnostic imaging ; metabolism ; pathology ; Male ; Middle Aged ; Platelet Endothelial Cell Adhesion Molecule-1 ; metabolism ; Radiography ; Sarcoma, Kaposi ; diagnostic imaging ; metabolism ; pathology ; Vimentin ; metabolism

OBJECTIVETo determine whether vitamin D receptor(VDR) gene start codon polymorphisms and 3'-end region polymorphisms exerted a combined influence on bone mineral density(BMD) in Han postmenopausal women in Beijing area.

METHODSThe VDR Fok I and 3'-end region genotypes were determined by polymerase chain reaction-restriction fragment length polymorphism in 110 unrelated postmenopausal women. BMD was measured at the lumbar spine (L(2-4)), femoral neck(Neck), Ward's triangle(Ward's) and trochanter (Troch) using duel-energy X-ray absorptiometry.

RESULTSThe frequencies distribution of Fok I, Apa I, Bsm I and Taq I alleles in this cohort all followed the Hardy-Weinberg equilibrium. No significant association of Fok I, Apa I or Taq I genotype with BMD in postmenopausal women was found when these polymorphisms were considered independently, except for Bsm I genotype. When a combined analysis of VDR gene Fok I and 3'-end region polymorphisms was carried out, cross-genotyping Fok I and Apa I polymorphisms was significantly associated with BMD at the L(2-4) (P<0.001), and cross-genotype of Fok I and Taq I was also significantly associated with BMD at the Neck and Troch sites (P<0.05). However, cross-genotyping Fok I and Bsm I polymorphisms was not significantly associated with BMD. Cross-genotyping Apa I and Bsm I or Taq I polymorphisms was not associated with BMD in postmenopausal women, either.

CONCLUSIONAlthough Fok I polymorphisms of VDR gene were not significantly associated with BMD in postmenopausal women, VDR gene Fok I and 3'-region polymorphisms (Apa I and Taq I) had a combined effect on the BMD in postmenopausal women.

3' Flanking Region ; genetics ; Aged ; Analysis of Variance ; Bone Density ; physiology ; China ; Codon, Initiator ; genetics ; DNA ; genetics ; metabolism ; DNA Restriction Enzymes ; metabolism ; Female ; Gene Frequency ; Genotype ; Humans ; Middle Aged ; Polymorphism, Genetic ; Polymorphism, Restriction Fragment Length ; Postmenopause ; genetics ; physiology ; Receptors, Calcitriol ; genetics