OBJECTIVETo compare the efficacy of different expression vectors, target genes, and immunization procedures in transfecting mice via liposome to construct murine model of Graves disease.

METHODSWe linked pCDNA3.1(+) and pUBC to full-length human TSHR and TSHR A subunit cDNA to yield four plasmids, which were later injected intramascularly or subcutaneously into female Balb/c mice via liposome. The blood anti-TSHR antibody (TRAb) were determined and the body weight were measured after each immunization. Serum thyroid hormone levels were measured after the animals were sacrificed.

RESULTSIn mice immunized with pUBC, no significant variance with control in weight nor serum TRAb concentration was observed. Weight gain in pCDNA3.1(+) group was significantlyly slower than controls (p<0.05), and serum TRAb concentration was also significantly elevated. In pCDNA group, animals immunized with TSHR A subunit (TSHRA subgroup) as the target gene revealed even significantly slower weight gain (p<0.001) and even faster TRAb elevation than those immunized with full length TSHR. Significantly higher FT4 (p=0.023) was observed in TSHRA and TSHR subgroups, which was reversely correlated to weight gain, but no significant difference (p>0.05) in FT3 was observed. Weight gain and TRAb concentration mainly varied in the later period of immunization.

CONCLUSIONSImmunization with pCDNA3.1(+) and TSHR A subunit gene together with higher immunization frequency increases the chance of model induction. Furthermore, FT4 is a better indicator for assessing the thyroid function in this model.

Animals ; Disease Models, Animal ; Female ; Graves Disease ; genetics ; Liposomes ; Mice ; Mice, Inbred BALB C ; Receptors, Thyrotropin ; genetics ; Transfection

AIMTo study on synthesis and antibacterial activity evaluation of polyheterocycles.

METHODSThe condensation of 4-amino-3-pyridin-3-yl-4H-[1,2,4] triazole-5-thiol with 2-chloromethyl-5-substituted phenyl-[1,3,4] oxadiazoles gave the corresponding title heterocycle amines, and the in vitro antibacterial activity was primarily evaluated by the method of cup-plate diffusion solution.

RESULTSTwelve novel compounds were synthesized, and their structures were confirmed by IR, 1H NMR, MS and element analysis. Biological screening results demonstrated that most of the compounds prepared showed good antibacterial activity.

CONCLUSIONOxadiazoles incorporting pyridyl triazole ring may be a pharmacophor structure in the molecule for developing antibacterial candidate drugs.

Anti-Bacterial Agents ; chemical synthesis ; chemistry ; pharmacology ; Escherichia coli ; drug effects ; Oxadiazoles ; chemical synthesis ; chemistry ; pharmacology ; Proteus vulgaris ; drug effects ; Staphylococcus aureus ; drug effects ; Triazoles ; chemical synthesis ; chemistry ; pharmacology

BACKGROUNDLidocaine and ropivacaine are often combined in clinical practice to obtain a rapid onset and a prolonged duration of action. However, the systemic toxicity of their mixture at different concentrations is unclear. This study aimed to compare the systemic toxicity of the mixture of ropivacaine and lidocaine at different concentrations when administered intravenously in rats.

METHODSForty-eight male Wistar rats were randomly divided into 4 groups (n = 12 each): 0.5% ropivacaine (group I); 1.0% ropivacaine and 1.0% lidocaine mixture (group II); 1.0% ropivacaine and 2.0% lidocaine mixture (group III); and 1.0% lidocaine (group IV). Local anesthetics were infused at a constant rate until cardiac arrest. Electrocardiogram, electroencephalogram and arterial blood pressure were continuously monitored. The onset of toxic manifestations (seizure, dysrhythmia, and cardiac arrest) was recorded, and then the doses of local anesthetics were calculated. Arterial blood samples were drawn for the determination of local anesthetics concentrations by high-performance liquid chromatography.

RESULTSThe onset of dysrhythmia was later significantly in group IV than in group I, group II, and group III (P < 0.01), but there was no significant difference in these groups (P > 0.05). The onset of seizure, cardiac arrest in group I ((9.2 + or - 1.0) min, (37.0 + or - 3.0) min) was similar to that in group II ((9.1 + or - 0.9) min, (35.0 + or - 4.0) min) (P > 0.05), but both were later in group III ((7.5 + or - 0.7) min, (28.0 + or - 3.0) min) (P < 0.05). The onset of each toxic manifestation was significantly later in group IV than in group I (P < 0.01). The plasma concentrations of the lidocaine-alone group at the onset of dysrhythmia (DYS), cardiac arrest (CA) ((41.2 + or - 6.8) min, (59.0 + or - 9.0) min) were higher than those of the ropivacaine alone group ((20.5 + or - 3.8) min, (38.0 + or - 8.0) min) (P < 0.05). The plasma concentrations of ropivacaine inducing toxic manifestation were not significantly different among groups I, II, and III (P > 0.05).

CONCLUSIONSThe systemic toxicity of the mixture of 1.0% ropivacaine and 2.0% lidocaine is the greatest while that of 1.0% lidocaine is the least. However, the systemic toxicity of the mixture of 1.0% ropivacaine and 1.0% lidocaine is similar to that of 0.5% ropivacaine alone.

Amides ; toxicity ; Anesthetics, Local ; toxicity ; Animals ; Arrhythmias, Cardiac ; chemically induced ; Cardiovascular System ; drug effects ; Central Nervous System ; drug effects ; Heart Arrest ; chemically induced ; Lidocaine ; toxicity ; Male ; Random Allocation ; Rats ; Rats, Wistar ; Seizures ; chemically induced

The purpose of this study was to investigate the growth characteristics and the expression level of integrin mRNA of the cultured bone marrow mesenchymal stem cells (BMMSCs) from patients with chronic myeloid leukemia (CML) in myeloid crisis (MC), and explore the role of BMMSCs in pathogenesis of CML. Five CML patients were enrolled in experimental group, five healthy persons were used as control. BMMSCs were cultured in vitro. The morphology of BMMSCs was observed every day and the growth curve were portrayed, and the ability of cell proliferation were detected according to the daily results of cell counting. Total RNA was extracted from third and fourth passages of BMMSCs, The expression of integrins mRNA of BMMSCs were measured by real-time PCR. The results showed that the BMMSCs of experimental and control groups had no difference in growth characterisctics, but the expression of integrins mRNA of the BMMSCs was higher in CML patients than in normal control group (p < 0.05). It is concluded that the abnormally high expression of integrins of BMMSC from the CML patients take part in pathogenesis of CML.
Adult ; Blast Crisis ; metabolism ; Bone Marrow Cells ; metabolism ; pathology ; Cell Proliferation ; Female ; Humans ; Integrins ; genetics ; metabolism ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; metabolism ; pathology ; Male ; Mesenchymal Stromal Cells ; metabolism ; pathology ; Middle Aged ; RNA, Messenger ; genetics ; metabolism ; Tumor Cells, Cultured

OBJECTIVETo identify factors associated with YMDD mutation in patients with chronic hepatitis B before and after lamivudine treatment in Zunyi region.

METHODS53 patients with chronic hepatitis B were enrolled in this study, HBV DNA,HBV markers, ALT, AST, TBil, albumin in the serum were examined at 0, 3, 6, 12, 18 and 24 months after lamivudine treatment. HBV genotype and YMDD mutation were determined by sequencing before lamivudine treatment. YMDD mutation was checked again if serum HBV DNA rebound to more than 1 x 10(4) copies/ml after the initial decrease.

RESULTSHBV genotype in Zunyi region is constitute of B, C and B+C genotype. YMDD mutation occurred in 18 cases after lamivudine treatment, the rate of YMDD mutation was 15.1%, and 34.0% after 1 year and 2 years treatment. There are four types of mutation: rtL180M/M204V, rtL180M/M204I, rtM204I, rtL180M. rtM204V mutation in C gene was always accompanied by rtL180M mutation (100%). The rate of rtL180M/M204V mutation in genotype C group was significantly higher than that in genotype B group (77.8% to 25.0%), the same was true for the rtL180M/ M204I mutation (22.2% to 12.5%). There was no point mutation in genotype C group. The point mutation of rtM204I and rtL180M appeared only in genotype B group. Gender, nation, family history of hepatitis B and HBeAg were not associated with YMDD mutation (P more than 0.05), while the mutation rate was associated with the disease course and severity of disease. YMDD mutation did not occur in patients with low HBV DNA level (less than 10(5) copies/ml).

CONCLUSIONYMDD mutation after lamivudine therapy is associated with HBV genotype and P gene mutation type, and prolonged treatment increases the the mutation rate. In order to reduce the incidence of YMDD mutation, patients with shorter disease course, lower HBV DNA level, more serious liver damage should be treated with lamivudine.

Adult ; Alanine Transaminase ; blood ; Antiviral Agents ; pharmacology ; therapeutic use ; Aspartate Aminotransferases ; blood ; China ; epidemiology ; DNA Mutational Analysis ; DNA Primers ; DNA, Viral ; blood ; genetics ; Drug Resistance, Viral ; Female ; Genotype ; Hepatitis B virus ; genetics ; Hepatitis B, Chronic ; blood ; drug therapy ; virology ; Humans ; Lamivudine ; pharmacology ; therapeutic use ; Male ; Mutation ; Polymerase Chain Reaction

Objective To explore the importance and methods of retaining articularis during pri- mary total hip arthroplasty(THA)and reconstruct soft tissue balance of hip joint after THA.Methods From February 2003 to August 2005,41 eases(43 hips)including 19 males and 22 females at age of 46- 80 years(mean 66.5 years)were treated with THA with retained capsule(Group R)and other 42 cases (44 hips)including 20 males and 22 females at age of 43-80 years(mean 64.3 years)with standard THA (Group S).Preoperative diagnosis found femoral neck fractures(GardenⅢⅣ)in 13 cases(13 hips)in Group R and 14(14 hips)in Group S;acetabular dysplasia(CroweⅢ)in 9(9 hips)in Group R and 8 (hips)in Group S;Osteoarthritis in 6(8 hips)in Group R and 7(8 hips)in Group S;and femoral head osteonecrosis(FicatⅢⅣ)in 13(13 hips)in Group R and 13(14 hips)in Group S.There were 13 hips of cement prostheses in Group R and 11 in Group S,8 cementless prostheses in Group R and 8 in Group S, 22 cement and cementless prostheses in Group R and 23 in Group S.Gibson's approach was used in both groups.Group R used the method of retaining capsule and little supination muscles during the operation to reconstruct responsibly soft tissue balancing of postoperation for THA.For comparison,Group S used the method of standard which resected a lots of capsule and didn't reconstruct it.The comparative items between Group R and Group S included incisional length,operative time,operative bleeding,drainage transfusion, infection,dislocation,postoperation standing,postoperation walking and Harris's score.Results All cases in Group R and Group S were followed for 6-22 months(mean 16.5 months in Group R and 16.7 months in Group S).There was significantly statistical difference upon interoperative and postoperative data between Group R and Group S.The result of Group R was significantly better than that of GS.Conclu- sion Retaining articularis during primary THA can minimize operative trauma,reconstruct soft tissue bal- ance and augment hip stability to get postoperative functional recovery.

This study examined the effect of Notch-1 signaling on malignant behaviors of breast cancer cells by regulating breast cancer stem cells (BCSCs). BCSCs were enriched by using serum-free medium and knocked out of Notch-1 by using a lentiviral vector. Real-time polymerase chain reaction (RT-PCR) and Western blotting were used to detect the Notch-1 expression levels in breast cancer cell lines and BCSCs, and flow cytometry to detect the proportion of BCSCs in BCSC spheres. The BCSC self-renewal, migration, invasion, and tumorigenicity were examined by the tumor microsphere-forming assay and transwell assay and after xenotransplantation. The results showed that the Notch-1 silencing reduced the number of BCSC spheres, the proportion of BCSCs, and the number of cells penetrating through the transwell membrane. It also decreased the size of tumors that were implanted in the nude mice. These results suggest that Notch-1 signaling is intimately linked to the behaviors of BCSCs. Blocking Notch-1 signaling can inhibit the malignant behaviors of BCSCs, which may provide a promising therapeutical approach for breast cancer.

Sorafenib is a multi -targeted tyrosinekinase inhibitor , and mainly used for the treatment of advanced renal cell carcinoma and hepa -tocellular carcinoma.The pharmacokinetic changes of sorafenib caused by drug -drug interactions may affect its clinical efficacy and safety, so individualized treatment drug monitoring could control the occurrence of potential drug interactions.In this paper, the clinical applications and drug -drug interaction of sorafenib was reviewed .

Objective To observe the clinical efficacy and safety of alendronate tablets combined with alfacalcidolon capsules in the treatment of perimenopausal women with osteoporosis.Methods A total of 56 perimenopausal women with osteoporosis were randomly divided into control group and treatment group with 28 cases per group.Control group was treated with alfacalcidolon capsule 0.25 μg qd oral.Treatment group was given alendronate sodium tablet 70 mg qd oral,on the basis of control group.The chnical efficacy,serum aquaporin 1 (AQP1),matrix metalloproteinase 2 (MMP-2),bone alkaline phosphatase (BALP),type Ⅰ collagen C end peptide (CTX-Ⅰ)and adverse drug reactions were compared between two groups.Results After treatment,the total effective rates in treatment and control groups were 92.86％ (26/28 cases) and 71.43％ (20/28 cases) with significant difference (P ＜0.05).After treatment,the main indexes in treatment and control groups were compared:AQP1 were (23.53 ± 3.02),(30.65 ± 4.31) μg · L-1;MMP-2 were (60.37 ± 8.35),(66.45 ± 8.47) μg · L-1;BALP were (20.04±2.57),(24.38 ±3.32) pg · L-1;CTX-Ⅰ were (0.75 ±0.06),(0.83 ±0.09)ng · L-1,the differences were statistically significant (P ＜ 0.05).The adverse drug reactions in treatment group were based on diarrhea and nausea,which in control group was based on diarrhea and nausea.The incidence of adverse drug reactions in two groups were 7.14％ without significant difference (P ＞ 0.05).Conclusion Alendronate tablets combined with alfacalcidolon capsules have a definitive clinical efficacy in the treatment of perimenopausal women with osteoporosis,which can significantly reduce the serum levels of AQP1,MMP-2,BALP and CTX-Ⅰ,without increasing the incidence of adverse drug reactions.

This study was undertaken to analyze and evaluate the diagnosis and principal treatment methods for congenital choledochal cyst, focusing on various surgical procedures and clinical outcome. A comprehensive, retrospective study was conducted on 72 adult patients who presented with choledochal cyst from 1985 to 2002. Surgical procedures were cyst excision with hepaticojejunostomy in 25 cases for type I or type IV-B, extrahepatic cyst excision with hepaticojejunostomy in 8 cases for type IV-A, extrahepatic cyst excision with modified hepaticojejunostomy in 2 cases for type IV-B, non-cyst excision with or without hepaticojejunostomy in 27 cases for types I, II, IV-A, IV-B. The early postoperative morbidity and mortality rate were 16.1% (9/62) and 6.5% (4/62) respectively, and the complication rate related to surgical procedure was 30.6% (19/62). The incidence of cholangiocarcinoma with non-cyst excision or non-operated congenital choledochal cyst was 10.8% (4/37). One patient died of primary hepatocellular carcinoma after cyst excision with hepatojejunostomy. In conclusion, our results showed that complete exci-sion of choledochal cyst for types I, II, and IV-B and complete excision of extra-hepatic choledochal cyst from the hepatic hilum in type IV-A with hepaticojejunostomy or modified hepaticojejunostomy are the treatment of choice for choledochal cyst in adult patients.
Academic Medical Centers/trends ; Adolescent ; Adult ; Choledochal Cyst/*epidemiology/*surgery ; Female ; Hepatectomy/*methods/*statistics & numerical data ; Humans ; Jejunostomy/*methods/*statistics & numerical data ; Korea/epidemiology ; Male ; Middle Aged ; Postoperative Complications/*epidemiology ; Retrospective Studies ; Treatment Outcome