To discuss the feasibility of the pharmacodynamic evaluation method for traditional Chinese medicine (TCM) formula particles, with traditional decoction for reference and the intervention of Magnoliae Officinalis Cortex in rats with ulcerative colitis (UC). First of all, the similarity of traditional Magnoliae Officinalis Cortex decoction and formula particles of different manufacturers was defined by using the IR fingerprint. The UC rat model was established and given Houpo formula particles of different doses and manufacturers, with the decoction for reference, in order to observe disease activity index (DAI), colon mucosa damage index (CMDI), pathologic changes, nitric oxide (NO), endothdin (ET), substance P, vasoactive intestinal peptide (VIP). Their intervention effects on UC rats were compared to study the difference between Sanjiu and Tianjiang Houpo formula particles, in order to demonstrate the feasibility of the pharmacodynamic evaluation method for Houpo formula particles. According to the results, Houpo formula particles showed similar pharmacodynamic actions with the traditional decoction. The pharmacodynamic comparison of Houpo formula particles of different manufacturers showed no statistical significance. The experiment showed that on the basis of the TCM compounds, a prescription dismantlement study was conducted to define target points of various drugs. The traditional decoction was selected for reference in the comparison of corresponding formula particles for their pharmacodynamic equivalence. This method could avoid controversies about single or combined boiling of formula particles, and give objective comments on the pharmacodynamic effect of the formula particles. The method is proved to be feasible.
Animals ; Chemistry, Pharmaceutical ; methods ; Colitis, Ulcerative ; drug therapy ; metabolism ; Dosage Forms ; Drug Evaluation, Preclinical ; Drugs, Chinese Herbal ; adverse effects ; chemistry ; pharmacokinetics ; Humans ; Magnolia ; chemistry ; Male ; Rats ; Rats, Wistar ; Substance P ; metabolism ; Vasoactive Intestinal Peptide ; metabolism

The knock out vector pHK was constructed with E coli vector pET 28a and shuttle vector pHY300PL, by using denatured DNA and homologous recombination technique, the kanamycin resistance gene ( Kan r) from integrated alkaline protease gene engineering strain BP071 was knocked out successfully, and the 11 positive clones were obtained The yield of the best positive clone BP0715 was stable as same as BP071 The methods provided the good experience for the industrial microbiology research, and it was foundation for studying on the safety of genetically modified organisms

Objective To clone lexA gene,express and purify the repressor LexA from Pseudomonas aeruginosa(PA),prepare the polyclonal antibody against PC-1 protein in rabbits,detect immunological activity of LexA protein.Methods The genomic DNA was extracted from the PAO1,the gene fragment encoding the mature LexA was amplied by PCR.It was linked the vector pET32a(+)and expressed in the E.coli BL21(DE3).The expressed protein was purified by two steps of Ni2+ chelate affinity chromatography and gel filtration chromatography respectively.The purified LexA protein immune the rabbits by injection and prepare the polyclonal antibody against PC-1 protein.The immunological activity of expressed and purified LexA protein was detected by ELISA,and Western blot.Results The expressed fused protein was found in insoluble form,accounted for 45% of the total bacteria protein.The final purity was 98.97%,which was determined by the HPLC.The expressed and purified LexA protein had satisfactory immunological activity.

Objective To investigate the clinical value of prenatal MRI in the diagnosis of fetal bowel obstruction.Methods Pregnant women suspected to have fetal abdominal abnormalities by ultrasonography were suggested to undergo MRI examinations within two days.Scanning sequence included FIESTA,SSFSE and T1WI SPGR sequence,with field of view focused on the fetal abdomen.After the final diagnoses of the cases were obtained by induced labor pathological examination or postpartum imaging or operation,the imaging data and the clinical data were reviewed and analyzed retrospectively.Results A total of 23 cases with bowel obstruction were included in the study.Four fetuses with duodenal atresia showed low T1 signal,high T2 signal characterized by double-bubble sign on MRI.There were 10 fetuses with jejunoileal atresia,showing bowel dilatation and hyperintense micro-colon on T1WI.Five cases of them depicted expansion of the terminal ileum with high T1 meconium signal.One each fetus had colonic atresia,intestinal malrotation with double-bubble and whirl sign.Annular pancreas with double-bubble sign and pressure trace of the bracket shape was detected in 3 fetuses.Meconium peritonitis was present in 4 fetuses,with 2 of them showing dilatation of intestine,ascites and pseudocysts.Conclusions According to the signal characteristics of amniotic fluid and meconium in the gastrointestinal tract on MRI,the obstructive level and development status of the distal bowel can be determined with MRI.It can provide additional information to ultrasonography,which brings clinical significance to prenatal diagnosis and intrapartum surgical operation.

Objective To evaluate the clinical applications and surgical methods of combined laparoscopic common bile duct (CBD) exploration with choledochoscopy. Methods From 2006 to 2009,clinical data of 42 patients with choledocholithiasis undergoing laparoscopic common bile duct exploration were retrospectively analyzed. We applied a step-by-step electric coagulating incision technique on the CBD,the step-by-step suturing technique, and the step-by-step clamping technique with alligator forceps, and soft tube irrigating technique with suctioning by selecting the proper exploration route, improving the common bile duct incision technique and calculus removing techniques. Results Procedures were successful in all the cases. There was no conversions to open surgery, no postoperative bleeding and no operative mortality. The mean operating time was 120 minutes (ranging, 90 to 150 minutes) with minimal intraoperative blood loss ( ranging, 20 to 40 ml). Ductal stone clearance was successful in 41 out of 42 patients ( 93% ). The largest number of the common bile duct stones was 16. With the diameter of stones larger than 15 mm in 18 cases in which the biggest was 30 mm. Bile leak developed in 1 patient, retained stones found in 3 patients,including intrahepatic cholelithiasis in one case. As a result, 38 out of 42 patients underwent common bile duct exploration. 35 patients were placed on T-tubes. Four patients underwent cystic duct exploration in which 3 had primary suture of the cystic duct and 1 had drainage. There was no infection and stenosis of biliary tract in the 42 followed-up cases. Conclusions Laparoscopic common bile duct exploration with stone extraction can be performed with high efficiency, minimal morbidity and without mortality. Improving the way of operation and selecting suitable exploration can result in better clinical outcomes.

OBJECTIVETo identify the genetic alterations in nonsyndromic cleft lip and palate (NSCLP).

METHODSComparative genomic hybridization was applied to investigate the genomic imbalance (the gain or loss of genetic material) in 7 cases of NSCLP.

RESULTSIt showed that the loss of chromosome DNA copies happened in chromosome 6, 7, 10, 13, 14, 16, 20, 22 and the gain of chromosome DNA copies happened in chromosome 5, 15, 18, 19. Conclusions 13q had a high frequency (71.4%) of chromosome loss.

CONCLUSIONSAbnormal chromosome DNA copies happen in all the patients with NSCLP. Most of the patients have chromosome DNA copies loss. It suggests that loss of inhibitory gene may be related to the NSCLP. The related inhibitory gene may locate in 13q.

Adolescent ; Adult ; Child ; Child, Preschool ; China ; ethnology ; Cleft Lip ; genetics ; Cleft Palate ; genetics ; Comparative Genomic Hybridization ; DNA ; Genetic Variation ; Genotype ; Humans ; Infant ; Mutation ; Phenotype ; Young Adult

Objective To understand the variation of thyroid hormones in each trimesters of pregnancy in women.Methods A total of 1 839 pregnant women in different trimesters were enrolled and thyroid hormones including FT3,FT4,TT3, TT4,TSH,TGAb and TPOAb were measured.The reference range of hormone in each trimester was calculated.Results There were statistically differences in all thyroid hormones among all trimesters of pregnancy women (P<0.05 )except for TT4 between the early and middle trimesters (P>0.05 ).The median of FT3 and FT4 were gradually reduced as the pregnant trimester grew up while the median of TSH had the adverse tendency.The median of TT3 was highest during the middle trimester (1.47 nmol/L).TT4 median was lowest during the late trimester (80.67 nmol/L).The TSH level of pregnant women over 30 years old was significantly lower than that of below 30 age group during all trimesters(P<0.05 ). There were significant differences in FT4,TT3 and TT4 level among early and middle trimesters among different ages groups(P<0.05 ).The positive rates of TGAb and TPOAb during late trimester (4.74% and 1.63%)were significantly lower than those of early trimester (1 4.40% and 5.56%)and middle trimester (1 4.40% and 5.56%)(P<0.05 ). Conclusion Significant differences of thyroid hormones are found not only in different stages of pregnancy,but also in different ages.Therefore,establishing trimester-specific reference data of thyroid hormones during different pregnency may be important for clinical practice.

OBJECTIVETo study the relation of the mRNA and protein expression of Cyclin A and p21cip1 in different stages hypertrophic scar fibroblast (FB) with its cell cycle, so as to provide theoretical evidence for intervention therapy of cell cycle gene being used in hypertrophic scar.

METHODSThe hypertrophic scar samples at different stages (the third month, the sixth month, the twelfth month, the twenty-fourth month) in 32 cases and the normal skin samples in 8 cases were used in this study. The expression of Cyclin A, p21cip1 mRNA and protein was detected by quantitative real-time PCR and Western Blot. And at the same time, the flow cytometer was used to detect the fibroblastic cell life cycle.

RESULTS1) The expression of Cyclin A mRNA and protein in the third month group, the sixth month group, the twelfth month group, the twenty-fourth month group were 19.34 +/- 2.41, 0.99 +/- 0.11; 19.30 +/- 1.42, 0.96 +/- 0.09; 10.73 +/- 2.93, 0.66 +/- 0.58; 9.29 +/- 0.97, 0.65 +/- 0.14, respectively. And in corresponding stages, the expression of p21cip1 mRNA and protein were 2.80 +/- 0.69, 0.35 +/- 0.07; 4.95 +/- 1.82, 0.44 +/- 0.07; 9.98 +/- 1.19, 0.56 +/- 0.06; 10.25 +/- 1.46, 0.59 +/- 0.06, respectively. The expression intensity of Cyclin A mRNA and protein was no significantly different between the third month group and the sixth month group (P > 0. 05), but it was higher respectively than that in the twelfth month group, the twenty-fourth month group and normal group (P < 0.05). And the expression intensity was no different between the above three groups (P > 0.05). The expression intensity of P21cip1 mRNA and protein in the third month group was lower than that in the sixth month group, but that in the above two groups was lower respectively than that in the twelfth month group, the twenty-fourth month group and normal group (P < 0.05). And the expression intensity had no difference between the three later stage groups (P > 0.05). 2) Most FB were in S and G2/M stage (cycle) in 3rd month, 6th month group. And most FB were in G0/G1 stage (cycle) in 12th month, 24th month group and normal group.

CONCLUSIONSThe expression of mRNA and protein of Cyclin A in hypertrophic scar changes from high level to low level as the hypertrophic scar develops, while the expression of P21cip1 changes from low level to high level. The mRNA and protein expression of Cyclin A and p21cip1 respectively are basically corresponded to different stages of hypertrophic scar. The distribution of cell life cycle of fibroblastic are also corresponded to the expression intensity of Cyclin A and p21cip1 in different stages hypertrophic scar. An early stage intervention to the two gene can be effective to prevent from the genesis and development of hypertrophic scar.

Adolescent ; Adult ; Cell Cycle ; Cells, Cultured ; Child ; Cicatrix, Hypertrophic ; metabolism ; pathology ; Cyclin A ; metabolism ; Cyclin-Dependent Kinase Inhibitor p21 ; metabolism ; Female ; Fibroblasts ; metabolism ; pathology ; Humans ; Male ; Middle Aged ; Young Adult

Biopsy ; Calcitriol ; therapeutic use ; Calcium ; therapeutic use ; Fanconi Syndrome ; diagnosis ; drug therapy ; Humans ; Immunosuppressive Agents ; therapeutic use ; Pilot Projects ; Prednisone ; therapeutic use

OBJECTIVETo explore the relationship between the mutations of epidermal growth factor receptor (EGFR) gene and clinicopathological characteristics in patients with non-small cell lung cancers (NSCLC).

METHODSParaffin-embedded tissue specimens were obtained from 1444 patients with NSCLC. The genomic DNA was extracted. Mutations of EGFR gene (exons 19 and 21) were detected by real-time PCR.

RESULTSDNA was available in 1410 cases. Somatic mutations of the EGFR gene were identified in 401 cases (27.8%). Among patients with EGFR mutations, 41.4% (n=166) had del E746-A750 of exon19, 6.7% (n=27) had del L747-P753insS of exon 19, 50.3% (n=201) had L858R of exon 21, and 1.5% (n=6) had L861Q of exon 21. Woman, non-smoker and adenocarcinoma showed a higher percentage of EGFR mutation (43.2%, 37.6%, and 33.5%, respectively). However, there was no association among age, grades, lymph node metastasis, and TNM stages (P>0.05). The mutation rate of BAC subtype (61.3%, 19/31) and adenocarcinoma with BAC features (48.0%, 12/25) was significantly higher than that of conventional adenocarcinoma (32.4%, 336/1038). A further assess of the smoking status found a trend that the more increased smoking exposure, the lower the incidence of EGFR mutations. A multivariable analysis revealed that adenocarcinoma, never smoking, and female were independently associated with EGFR mutations (odds rations=3.381, 2.393, and 1.727, respectively).

CONCLUSIONSThe detection rate of EGFR mutation is higher in Chinese patients, especially in non-smoking female patients with adenocarcinoma. Real-time PCR is a sensitive and accurate method to detect the mutations of EGFR gene and can therefore provide useful information for clinical treatment.

Adenocarcinoma ; genetics ; Adenocarcinoma, Bronchiolo-Alveolar ; genetics ; Adult ; Aged ; Aged, 80 and over ; Carcinoma, Non-Small-Cell Lung ; genetics ; pathology ; Exons ; Female ; Genes, erbB-1 ; genetics ; Humans ; Lung Neoplasms ; genetics ; pathology ; Male ; Middle Aged ; Mutation ; Mutation Rate ; Real-Time Polymerase Chain Reaction ; Receptor, Epidermal Growth Factor ; genetics ; Sex Factors ; Smoking ; Young Adult