Fluorescence enhancement reaction of flavoxate hydrochloride (FX) in strong alkali solution was studied, the mechanism of the reaction was investigated, and a novel fluorimetric method for analysis of FX in drug sample was established. FX has no intrinsic fluorescence, but it can slowly produce fluorescence in strong alkali solution. Heating can promote the fluorescence enhancement reaction. In 3D fluorescence spectra of the decomposition product of FX, two fluorescence peaks, located respectively at excitation wavelengths λex/ emission wavelength λem =223/410 nm, and 302/410 nm, were observed. Using quinine sulfate as a reference, fluorescence quantum yield of the decomposition product was measured to be 0.50. The structural characteriza- tion and spectral analysis of the decomposition product reveal that ester bond hydrolysis reaction of FX is firstly occurred during heating process, forming 3-methylflavone-8-carboxylic acid (MFA), then a cleavage reaction of the γ-pyrone ring of MFA occurred, producing α, β-unsaturated ketone. This product includes adjacent hydroxyl benzoic acid group in its molecule, which can form intramolecular hydrogen bond under alkaline condition, so that increase the conjugate degree and enhance the rigidity of the molecule, and thereby cause fluorescence enhancement. Based on this fluorescence enhancement reaction, a fluorimetric method was proposed for the determination of FX. A linear calibration curve covered the concentration range 0.020 3-0.487 µg · mL. The regression equation was I(F) = 23.9 + 5357.3 c, with correlation coefficient r = 0.999 7 (n = 8), detection limit D = 1.1 ng · mL(-1). The method was applied to the analysis of FX tablets, with a spiked recovery rate of 100.2%. The reliability of the method was verified by a UV-spectrophotometric method.
Alkalies ; Calibration ; Chemistry, Pharmaceutical ; Flavoxate ; analogs & derivatives ; chemistry ; Fluorescence ; Limit of Detection ; Reproducibility of Results ; Solutions ; Tablets

Three-dimensional (3D) fluorescence spectra and thin layer fluorescence chromatogram of common Camptotheca fruit (CCF) crude drug, camptothecin (CPT) and 10-hydroxycamptothecin (HCPT) have been studied, and a novel fluorimetric method for determination of CPT in CCF crude drug has been established. In 3D fluorescence spectra, CPT presented 3 fluorescence peaks with excitation wavelengths lambdaex of 215, 255 and 365 nm, separately, and all peaks with emission wavelength lambdaem of 430 nm. HCPT presented 4 fluorescence peaks with lambdaex of 220, 265, 325 and 375 nm, separately, and all peaks with lambdaem of 555 nm. The fluorescence of CPT is much stronger than that of HCPT. Comparison of 3D fluorescence spectra and analysis of thin layer fluorescence chromatogram revealed that the main fluorescent component of CCF is CPT. HCPT and other components basically do not interfere with the fluorescence of CPT. Under the condition of pH 3.0-6.7, CCF aqueous solution can produce strong and steady fluorescence. Using methanol as solvent, the extracting solution of CCF was prepared, and diluted properly with water, then measured fluorescence intensity at lambdaex/lambdaem = 365/430 nm to determine the content of CPT. A linear calibration curve covered the concentration range 0.002 35-0.235 microg x mL(-1). The regression equation was IF = 9 + 30,844 c, with correlation coefficient r = 0.999 (n=9). The method has been applied to the analysis of CPT in a CCF sample, with a result of 0.127% and a spiked recovery rate of 102%. The reliability of the method has been verified by a thin layer chromatography-fluorescence scanning method. Experimental results demonstrated that this method can be used for quality evaluation of CCF crude drug.
Antineoplastic Agents, Phytogenic ; analysis ; Camptotheca ; chemistry ; Camptothecin ; analogs & derivatives ; analysis ; Chromatography, Thin Layer ; Fruit ; chemistry ; Plants, Medicinal ; chemistry ; Quality Control ; Reproducibility of Results ; Spectrometry, Fluorescence

Adenoviridae ; genetics ; Calcium-Binding Proteins ; genetics ; metabolism ; Cells, Cultured ; Female ; Genetic Vectors ; Humans ; Microfilament Proteins ; genetics ; metabolism ; Myocytes, Smooth Muscle ; metabolism ; Myometrium ; cytology ; metabolism ; RNA, Small Interfering ; genetics ; Recombinant Fusion Proteins ; genetics ; metabolism ; Transfection

Adult ; Carcinoma, Renal Cell ; secondary ; Female ; Humans ; Kidney Neoplasms ; pathology ; Magnetic Resonance Imaging ; Vaginal Neoplasms ; secondary

Objective:To produce rhBMP-4 with bioactivity in E.coli. Methods: The full-length human BMP-4 gene was mutated by PCR without changes in amino acid sequence, then the synthesized gene was cloned into plasmid pET-3c, transducted into BL21(DE)plysS, and induced by adding IPTG to a final concentration of 1.0 mmol/L. The protein product was purified using ion-exchange chromatography method and then renaturated, bioactivity was checked by C2C12 differentiation in vitro and mouse ectopic bone formation in vivo. Results: A 438 bp gene fragment encoding mature peptide of hBMP-4 was cloned , the protein product was mostly in the form of inclusion body, after renaturation, the engineering protein shows better bioactivity. Conclusion:The mutant strategy can enhance the expression of bioactive rhBMP-4 in E.coli expression system.

OBJECTIVETo evaluate the expression of microRNAs and reveal the regulatory mechanism of miRNA-708 in pediatric common acute lymphoblastic leukemia (ALL) (common-ALL).

METHODSThe expressions of microRNAs in common-ALL patients were detected by microarrays in 3 pediatric common-ALL samples, and then verified by stem-loop quantitative RT-PCR in 34 common-ALL samples. The target genes of miR-708 were found by bioinformatics software, and verified by dual-luciferases reporter assay, RT-PCR and Western blot.

RESULTSCompared to normal bone marrow samples, of all the 2006 detected miRNAs, the expression of miR-708, miR-181b and miR-210 were 16.886 ± 16.854, 5.710 ± 4.652, and 9.789 ± 1.178, retrospectively, being significantly up-regulated expressed than those in normal control (1.872 ± 0.339, 1.276 ± 0.531 and 1.005 ± 0.080, retrospectively) (P < 0.05), while miR-27b and miR-345 were the two most down-regulated ones (0.524 ± 0.085 and 0.675 ± 0.086, retrospectively) (normal control: 1.123 ± 0.066 and 1.204 ± 0.140, retrospectively) (P < 0.05). And the expression of miR-708 and miR-181b were significantly correlated with the clinical types in common-ALL. In high risk common-ALL, miR-708 and miR-181b were much higher than in standard and middle risk common-ALL (P < 0.05). The further verification research in 293 cell line showed that miR-708 decreased the expression level of its target genes CNTFR, NNAT and GNG12 by combining with 3'-UTR of the 3 genes, moreover, miR-708 combined with CNTFR 3'-UTR in 394 ∼ 400 bp sequence region.

CONCLUSIONMicroRNAs plays an important regulatory role during the occurrence and development of the pediatric common-ALL and miR-708 is an important factor for high risk common-ALL.

Child ; Child, Preschool ; Female ; Gene Expression Profiling ; Humans ; Infant ; Male ; MicroRNAs ; genetics ; metabolism ; Microarray Analysis ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; genetics

OBJECTIVETo introduce statistical methods of time trend analysis on cancer rates.

METHODSCancer incidence data collected by the Shanghai Cancer Registry during 1991 to 1999 was used in the analysis to calculate the crude and age-adjusted rates, percent changes (PCs) and annual percent changes (APCs). APCs were estimated by a linear regression of the logarithm on the incidence rates during the nine years. It also introduced a method for partitioning a linear trend in age-adjusted rates into site-specific contributions to the overall floating trend. 95% confidence intervals for the APCs and contributions were described in the paper.

RESULTSA decreasing rates were observed for cancers of stomach and esophagus among both men and women in urban Shanghai from 1991 to 1999. The increasing rates among men would include cancers of colon, rectum, gall bladder, pancreas, prostate, urinary bladder, kidney and leukemia. The rates of cancers among women increased for colon, rectum, lung, breast, gall bladder, endometrium, ovary, urinary bladder and kidney. The changes of above cancers over time were statistically significant (P < 0.05 or P < 0.01), but rates for other cancer sites changed little. The APCs (weighted method) and contributions for the cancers of stomach, esophagus, colon, rectum and prostate were -2.99% and -65.72%, -2.90% and -17.07%, 12.30% and 21.46%, 2.94% and 18.62%, and 3.11% and 15.09% among men, and -6.05% and -39.55%, -1.08% and -35.19%, 2.81% and 28.64%, and 3.69% and 15.70% for the cancers of stomach, esophagus, breast and colon in women, respectively.

CONCLUSIONAPC, and related statistics could be used to describe and analyze the time trend of cancer rates rather than PC or/and graphical method alone.

Algorithms ; China ; epidemiology ; Female ; Humans ; Incidence ; Linear Models ; Male ; Neoplasms ; epidemiology ; Time Factors

OBJECTIVETo explore the norms of treatment of acute organophosphorus pesticide poisoning (AOPP), and observe the curative effect.

METHODSOn basis of the pre-research, the norms of treatment of AOPP were summarized, and a multi-center clinical trial was performed in 6 hospitals selected from high incidence of AOPP in Shandong Province.

RESULTS422 patients of AOPP in 6 hospitals in observation period were treated and observed by the norms of treatment. Among them, the proportion of oral poisoning was 97.16%, middle and severe degree were 87.44%. Compared with themselves 2 years ago before standard treatment, the curative effect of the norms of treatment for AOPP was much better than before. The mortality rate of AOPP declined from 9.87% to 1.66% (Chi2 = 27.92, P < 0.01), that was much better than the average therapeutic effect level of all our province in the same period (the mortality rate: 8.92%) (Chi2 = 26.05, P < 0.01). The average amount of atropine [(37.54 +/- 17.76) mg], dropped greatly [(1280.70 +/- 69.22) mg] (U = 439.22, P < 0.01).The usage of atropine by continuous intravenous injection with venous pump was better than ordinary intravenous injection. The mean dosage of pralidoxime chloride increased twice than the previous (U = 19.48, P < 0.01). There was no drug poisoning.

CONCLUSIONThe standard treatment of AOPP is urgently needed in our country, especially in rural area. By this trial, the satisfactory effect of the norms of treatment for AOPP summarized is observed and it reduces the fatality rate remarkably.

Adolescent ; Adult ; Aged ; Child ; Female ; Humans ; Male ; Middle Aged ; Organophosphate Poisoning ; Pesticides ; poisoning ; Poisoning ; therapy ; Standard of Care ; standards ; Young Adult

Objective Clinical data of 19 Chinese patients with 21 hydroxylase deficiency (21OHD) were analyzed to improve the diagnosis and treatment level. Methods Clinical features and laboratory data were collected from 19 patients with 21OHD before and after treatment. Results In male patients, the average age of early appearance of secondary sexual character was (9.3?2.8)yrs, and excess androgen resulted in phallic enlargement. Primary amenorrhea was the most common complaint in female(87.5%), and the signs included a varying degree of labioscrotal fusion and clitoral enlargement. The average level of 17-hydroxy progesterone(17OHP) was (63.42?35.07) ?g/L, and adrenocorticotrophic hormone(ACTH), dehydroepiandrosterone(sodium) sulfate(DHEAS) and testosterone(T) were obviously elevated. CT scan showed bilateral adrenal hyperplasia. The level of 17OHP was significantly decreased after treatment[(63.42?35.07) ?g/L vs (3.15?2.71) ?g/L](P

OBJECTIVETo evaluate the early and mid-term outcome of surgical repair for post-ductal coarctation of the aorta (CoA) under normothermia without cardiopulmonary bypass.

METHODSClinical data from 15 patients (11 males, 4 females, mean age 18 +/- 10 years) undergoing surgical repair for post-ductal CoA under normothermia without cardiopulmonary bypass between January 1999 and December 2004 were analyzed retrospectively. There were 7 isolated cases, 7 cases associated with patent ductus arterious (PDA), 1 case with PDA and ventricular septal defects. Operation was performed under normothermia with partial cross-clamping of descending aorta in 8 cases, compete cross-clamping in 6 cases and temporary shunt in 1 case. Operative techniques adopted prosthetic bypass graft in 9 cases, Gore-Tex patch graft aortoplasty in 4 cases and stenosis resection with end-to-end anastomosis in 2 cases. PDA was ligated at single-stage in 8 cases. Ventricular septal defect was repaired at second stage in 1 case.

RESULTSNo early and late death. Hypertension occurred in 9 cases during early postoperative period but was normalized gradually in 5 cases without medication during follow-up period, from 6 months to 5 years. The arterial blood pressure of lower extremities increased significantly and no hoarseness, paraplegia occurred after operation. No recoarctation and aneurysm formation were found during follow-up.

CONCLUSIONSurgical repair of post-ductal CoA under normothermia without cardiopulmonary bypass is safe and effective, which is a procedure of choice for patients with isolated CoA, CoA associated with PDA, or with other intracardiac anomalies that are ready to be repaired at second-stage.

Adolescent ; Adult ; Aortic Coarctation ; surgery ; Cardiovascular Surgical Procedures ; methods ; Child ; Child, Preschool ; Female ; Follow-Up Studies ; Humans ; Male ; Retrospective Studies ; Temperature ; Treatment Outcome