ObjectiveTo observe the preventative and therapeutical effect on bile pigment calculus of Jinqianduizuo Mixture which is a traditional Chinese medicine empirical formula and provide foundation for the therapy of cholelithiasis. MethodUsing the model of bile pigment calculus in guinea pigs. Observing the gall bladder’s shape,the rate of forming gallstone and the total bile. Then making biochemical determination of the bile and the blood serum. Result There are significant differences between model control group and three treated groups of Jinqianduizuo Mixture in the rate of forming gallstone,the bilious quantity,the contents of total bilirubin and total bile acids. While in the contents of direct bilirubin,total cholesterol,triglycerides and high-density lipoproteins,they have no significant difference. ConclusionThe results indicate that Jinqianduizuo Mixture can conspicuously cut down the gallstone’s incidence of guinea pigs and can obviously promote the bilious externalization in guinea pigs. It also can promote bile secretion distinctly,but have no evident effect on the ingredients of blood-lipid.

[Objective] To discuss mechanism of FS-1272 eye drops on cataract of rats induced by D-galactose.[Method]By using the model of cataract of rats induced by D-galactose to determine the content of SOD,CAT,GSH-px,MDA in lens.[Result]FS-1272 eye drops can increase the activities of SOD,CAT,GSH-px and decrease the content of MDA.[Conclusion]FS-1272 eye drops have the effect of delaying the generation and development of cataract by alleviating the extent of oxidative injury of lens.

[Objective] To elucidate the effects and mechanism of Naoqingling Decoction(ND)on Gilles de la Tourette syndrome(TS).[Method] Central nervous system of mice was stimulated by amphetamine(AMP)to develop the model of experimental tourette syndrome(ETS).Then we observed the stereotypia and autonomic activities of mice,and determined the levels of monoamine neurotransmitters such as dopamine(DA),norepinephrine(NE),5-hydroxytryptamine(5-HT)and 5-hydroxyindoleacetic acid(5-HIAA)in the brains.[Result] To compare with model group,three-dose ND groups can resist the determinate activities stimulated by AMP;can obviously decrease the autonomic activities of mice and degrade the levels of DA,NE,5-HT and 5-HIAA in the corpora striates.[Conclusion] ND has therapeutical effect on ETS model and it's mechanism may be its effects on the secretion or metabolism of neurotransmitter in the brains.

A 65-year-old man with right central type of lung squamous carcinoma was admitted to our department. Bronchoscopy displayed complete obstruction of right upper lobe bronchus and inifltration of the bronchus intermedius with tumor. Chest contrast computed tomography revealed the tumor invaded right pulmonary artery, superior vena cava, and the persistant letf superior vena cava lfowed into the coronary sinus. hTe tumor was successfully removed by means of bronchial and pulmonary artery sleeve resection of the right upper and middle lobes combined with resection and reconstruction of superior vena cava (SVC) utilizing ringed polytetralfuoroethylene gratf. To the best of our knowledge, this was the ifrst report of complete resection of locally advanced lung cancer involving superior vena cava, right pulmonary artery trunk and main bronchus with persistant letf superior vena cava.

Objective: To describe the prevalence of permanent tooth caries and associated factors among junior high school students in Haikou, and to provide reliable evidence for prevention and control of caries. Methods: A total of 3 573 students from 8 junior high school in Haikou City and towns were selected by the methods of clustered sampling survey. Questionnaire survey and oral health examinations were conducted to analyze the caries rate, mean decayed missing filled tooth (DMFT), filling rate, pit and fissure sealant rate. Logistic regression was used to analyze the influencing factors of caries in permanent tooth. Results: Among the surveyed junior high school students, the caries rate was 47.32%( n =1 691), the mean DMFT was 1.51. The caries rate and mean DMFT were higher in female students (49.59%) than in male students (44.95%), and higher in towns (50.77%) than in urban areas (44.04%), the difference were statistically significant ( χ 2=16.25, 7.72, P <0.05). The filling rate and pit and fissure sealant rate among junior high school students in Haikou were 17.13%, 6.27% respectively. The filling rate and pit and fissure sealant rate were higher in urban areas (18.97%, 7.17%) than towns (15.76%, 5.33%), the difference were statistically significant ( χ 2= 9.57, 5.13, P <0.05). The multivariate Logistic analysis showed that female student, town residence, daily consumption of sweets or sugary drinks (≥1 time), bedtime snack were risk factors for junior high school students suffering from permanent dental caries( OR =1.41, 1.45, 2.63, 2.09, 1.53), while using fluoride toothpaste daily, oral education in school were protective factors( OR =0.44, 0.34)( P <0.05). Conclusion The caries rate of permanent teeth among junior high school students in Haikou is at high level, but the filling rate of caries and pit and fissure sealant rate are lower. The prevention and treatment of dental caries should be carried out in high risk students, and oral health education in school is also needed to improve the oral health level of junior high school students.

A single-stranded oligonucleotides containing a 6 histidine sequence, a hydroxylamine cleavage site, a thrombin cleavage site, and stop codon TAA were inserted into the polylinker's downstream of prokaryotic expression vector pBV220 between BamHI and PstI. The resultant vector is named pBV223. Proteins expressed in this vector will have a 6 histidine tail as affinity handy fused to their C terminus and can be quickly purified by one step immobilized metal affinity chromatography (IMAC). This plasmid is verified by restriction map and DNA sequencing. Subsequently, the metastasis suppressor gene nm23-H1 cDNA (without the stop codon) was cloned into vector PBV223 in frame with the 6-histidine sequence, hydroxylamine and thrombin cleavage sites. The soluble nm23-H1 fusion protein was successfully induced in the bacterial DH5a and easily purified with Ni chromatograph. These results indicated that the strategy to clone the single-stranded oligonucleotides directly into the restriction sties between BamH I and Pst I in the pBV220 vector is the simplest and cost-effective method.
Base Sequence ; Chromatography, Affinity ; methods ; Cloning, Molecular ; DNA, Complementary ; genetics ; Escherichia coli ; genetics ; metabolism ; Genetic Vectors ; genetics ; Molecular Sequence Data ; Mutation ; NM23 Nucleoside Diphosphate Kinases ; biosynthesis ; genetics ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; isolation & purification

BACKGROUNDTo summarize the clinical results of bronchoplastic procedures and pulmonary artery reconstruction or combined with other resection and plasty of heart, great vessels in the treatment of 304 patients with locally advanced lung cancer.

METHODSFrom February, 1983 to December, 2001, double sleeve resection and reconstruction of bronchus and pulmonary artery, or combined with other resection of heart, great vessels were carried out in 304 patients with locally advanced lung cancer. The operations included double sleeve left upper lobectomy in 199 cases; double sleeve right upper lobectomy in 21 cases; double sleeve right upper middle lobectomy in 14 cases; double sleeve left upper lobectomy combined with resection of left atrium in 8 cases; double sleeve right upper lobectomy combined with superior vena cava (SVC) resection and reconstruction with Gortex graft in 29 cases; double sleeve right upper middle lobectomy combined with SVC resection and reconstruction in 21 cases; double sleeve right upper middle lobectomy, carinal and SVC resection and reconstruction in 11 cases; left pneumonectomy combined right main pulmonary artery and pulmonary artery trunk resection and reconstruction with Gortex graft in 1 case.

RESULTSThere were 3 operative deaths. The operative mortality was 1% in this series. Sixty four patients had operative complications. The operative complication rate was 21.05% (64/304). The 1-, 3-, 5- and 10 year survival rates were 81.75%, 60.14%, 37.21% and 24.39% respectively.

CONCLUSIONSDouble sleeve lobectomy or comblined with other resection and reconstruction of heart, great vessels can significantly improve the prognosis and increase the curative rate and long term survival in patients with locally advanced lung cancer.

BACKGROUNDThe present experimental data have showed that the function of kinase suppressor of Ras (KSR) is mainly as a scaffold protein that coordinates the assembly of a multiprotein complex containing MAPK and its upstream regulators. But whether KSR has kinase activity is still the point of argument until now. The aim of this study is to construct eukaryotic expression vectors of carboxyl terminus and amino terminus of KSR and to detect their expression in 293T cell line.

METHODSN-KSR and C-KSR were amplified by PCR. The eukaryotic expression vectors of pCMV-Tag2b-N-KSR and pCMV-Tag2b-C-KSR were constructed by gene recombination technique and the recombinant plasmids were verified by restriction enzyme analysis and sequencing. Then positive clones were transfected into 293T cell line. Expression of target proteins was analyzed by Western blot.

RESULTSThe sequences and open read frames of the two vectors were both completely concordant with experiment design. The target proteins could be observed in transfected 293T cells by Western blot.

CONCLUSIONSEukaryotic expression vectors of pCMV-Tag2b-N-KSR and pCMV-Tag2b-C-KSR are successfully constructed, and they can be expressed in 293T cells. It provides an experimental base for further research work.

BACKGROUNDPrevious researches have proven that nm23-H1 gene is a tumor metastatic suppressor gene, however, it is still unknown about its exact molecular mechanisms. Site-directed mutagenesis can correctly change the base sequence and get mutant proteins. The aim of this study is to construct nm23-H1 mutant and EGFP fusion genes by site-directed mutagenesis, and to provide basement for studying the functional and biochemical mechanisms of nm23-H1 gene.

METHODSSite-directed mutagenesis of nm23-H1 gene was performed by modified QuikChange™ Site-directed Mutagenesis Kit method. Pure plasmid containing fusion gene of nm23-H1 and EGFP (PLXSN-nm23-H1-EGFP) was mini-prepared. Four pairs of mutagenic primers were synthesized in vitro and the desired five mutations, S44A, P96S, H118F, S120G and P96S-S120G were introduced into nm23-H1-EGFP fusion gene by PCR.

RESULTSFive nm23-H1 mutant and EGFP fusion genes, nm23-H1(S44A)-EGFP, nm23-H1(P96S)-EGFP, nm23-H1 (H118F)-EGFP, nm23-H1(S120G)-EGFP and nm23-H1(P96S-S120G)-EGFP, were successfully constructed. The results of DNA sequencing confirmed that the base sequences of the mutant fusion genes were completely concordant with experiment design.

CONCLUSIONSFive nm23-H1 mutant and EGFP fusion genes are successfully constructed, which can be used in further studies. QuikChange™ site-directed mutagenesis is a simple, rapid and efficient method.

BACKGROUNDIt has been proved that lysosome-associated protein transmembrane 4 beta (LAPTM4B) plays an important role in tumorigenesis, the *2/*2 genotype of LAPTM4B was closely related to liver cancer susceptibity. The aim of this study is to investigate the possible association between the allelic variation of LAPTM4B and the genetic susceptibility of lung cancer.

METHODSThe genotype of LAPTM4B was detected in 131 patients with lung cancer and 104 unrelated healthy adult individuals as control by polymerase chain reaction based on special primers. The genotypic distribution of LAPTM4B was analyzed by Chi-square test.

RESULTSThe allelic frequencies of the *1 and *2 were 74.8% and 25.2% in the lung cancer group and 72.1% and 27.9% in the healthy control group respectively, but no significant difference was observed between the two groups (Chi-square=0.433, P=0.510). The genotypic distribution of the *1/*1,*1/*2 and *2/*2 were 53.4%, 42.7% and 3.8% in the lung cancer group, 54.8%, 34.6% and 10.6% in the healthy control group respectively, there was no significant difference between the two groups (Chi-square=4.89, P=0.087). No remarkable association was observed between the genotypic distribution of LAPTM4B and the clinical information in the patients with lung cancer, including pathological type and TNM staging.

CONCLUSIONSThe results suggest that the allele of LAPTM4B is not closely associated with genetic susceptibility of lung cancer.