This article reviews the resultsof the basic research about epidermal growth factor and its receptor,and the development of the novel drug, EGF eyedrop, that containing chemically synthesised EGF gene,the construction of EGF expression vector, the transformation of the host cells , the purification of the recombinant protein EGF, the preparation of three batches of the EGF product and identification, the preclinical and clinical trials. Relevant studies show that recombinant EGF consisting of 51 amino acids can be secreted into the medium under the control of the α factor leading sequence in the yeast cells.The EGF can accelerate the growth of corneal-limbal epithelial cells and the healing of an alkali burned corneal. The EGF can be used in curing oral cavity ulcer and skin burned wound. And it has the preventive effects on experimental duodenal ulcer of rat. The antiserum was made for test of the con- centration of blood EGF and urine EGF by RIA. Data from studies demonstrate the inhibition effect of EGF on the growth of tumor cells, such as A431 and BT325 cells in the presence of high EGF concentration (＞10 ng/ml). The expression of EGFR and DNA ploidy in renal carcinoma has clinical significance. Crystallization and preliminary x-ray diffraction studies of the EGF has been made. The MW of the EGF product is 6000, and the pI is about 4. 6 and it has correct N-terminal amino acids sequences , immunogenicity and biological activity. There is no vestige of the DNA of the yeast cells. Animal experiments reveal that there is no cumulation of the EGF in the body, and EGF can promote corneal epithelial healing. There is no toxicological effect during cornea wound healing of rabbit. A randomized, double-blind, placebo-controlled, multi-center clinical trial was conducted in four hospitals to assess safety,ocular tolerance and efficacy of an ophthalmic solution of EGF for 200 cases of cornea transplantation and 247 cases of nebulae. Unequivocal results were obtained as the eyedrop really accelerate the wounded cornea healing. So, the EGF eyedrop as a novel drug of class I is approved by the National Drug Administration, and this is the first gene engineering drug that come from yeast expression system in China.

Objective To examine the complement component 1 Q subcomponent-binding protein (C1QBP) gene expression in human resistance choriocarcinoma cell lines and its parental cell line JeG-3,and to investigate whether silence C 1QBP by small interference RNA could reverse the resistance of human resistance choriocarcinoma cell lines to its relevant chemotherapy drugs.Methods Expression of C1QBP mRNA and protein in cells were detected by real-time fluorogenic quantitative PCR and western blot,respectively.The difference of C 1QBP expression was compared between human resistance choriocarcinoma cell lines and its parental cell line JeG-3.Sub-cellular location was proved by confocal immunofluorescence microscopy.A lentiviral vector containing short hairpin RNA (shRNA) targeting C 1QBP was constructed and cotransfected with the packaging plasmid mixture into 293T cells by lipofectamine 2000.The human resistance choriocarcinoma cell lines were infected with the packaged lentivirus.Real-time fluorogenic quantitative PCR and western blot were used to validate whether the C 1QBP gene expression was silenced.The cell counting kit 8(CCK8)was used to determine the drug sensitivity.Results (1)The C1QBP mRNA expression levels among four human resistance choriocarcinoma cell lines[JeG-3/floxuridiuum (FUDR),JeG-3/methotrexate (MTX),JeG-3/etoposide (VP),JeG-3/dactinomycin (KSM)] were 2.520±0.680,1.770±0.230,1.940±0.090 and 1.740±0.350 folds compared to that in JeG-3 cells.The C1QBP protein was higher expression level in human resistance choriocarcinoma cell lines than that in JeG-3.The immunofluorescence methods and confocal analysis showed that C1QBP localized predominantly in the mitochondrial matrix.(2)The C1QBP mRNA expression in JeG-3/FUDR cells after infected with lentiviral vector were decreased by 93.1％ (P＜0.01).The protein expression of C 1QBP in JeG-3/FUDR cells after infected with lentiviral vector were almost completely suppressed.The resistance indexes of four human resistance choriocarcinoma cell lines(JeG-3/FUDR,JeG-3/MTX,JeG-3/VP,JeG-3/KSM) were respectively 86.3％,93.9％,92.8％ and 89.9％,which were decreased remarkably by knockdown the C 1QBP expression (P＜0.05).Conclusions C1QBP is overexpressed in human resistance choriocarcinoma cell lines compared with parental cell line JeG-3.Inhibition of C 1QBP by lentivirus-mediated small interference RNA could effectively reverses the resistance of human resistance choriocarcinoma cell lines to its relevant chemotherapy drugs.

Objective To investigate the clinical effect of porcelain fused for metal crown restoration after root canal therapy.Methods Totally 90 patients with pulpitis or periapical periodontitis from December 2013 to December 2015 had their clinical data analyzed,who underwent root canal therapy and were divided equally into an observation group and a control group.The observation group took porcelain endocrowns repair and the control group applied ceramic crown.The two groups were compared on the restoration integrity,edge sealing,residual rate,gum,food impaction and color 3 months,6 months and 1 a after treatment.Results The observation group behaved significantly better than the control group in the edge sealing and restoration integrity (P＜0.05).There were significant differences between the residual rates,gum and food impaction in the two groups (P＜0.05).Conclusion Porcelain endocrowns repair gains advantages over ceramic crown repair in edge sealing,integrity,gum,food impaction and color after root canal therapy,though its long-term effect needs further investigation.

AIMTo apply stochastic resonance algorithm (SRA) to quantitative analysis of weak chromatographic signal, which was embedded in the noise.

METHODSBased on the theory of stochastic resonance (SR), a simple and effective SRA has been established to improve analytical detection limits of chromatographic analysis, which apply to enhance the signal to noise ratio by the optimization of the parameters and Runge-Kutta method, was established. The method was used to quantitative analysis of phenazopyridine in human plasma by HPLC/UV. Meanwhile this method is compared with HPLC/MS.

RESULTSBy experimental chromatographic data sets, an excellent quantitative relationship between concentrations of phenazopyridine and their responses had been obtained. The concentration of phenazopyridine in plasma determined by HPLC/UV with SRA and HPLC/MS showed that there was no significant difference (P > 0.05) between the two methods.

CONCLUSIONThe new method was feasible.

Chromatography, High Pressure Liquid ; methods ; Humans ; Mass Spectrometry ; Noise ; Phenazopyridine ; blood ; Sensory Thresholds ; Spectrophotometry, Ultraviolet ; Stochastic Processes

To investigate the antitumor mechanism of artemisninin, a flexible docking analysis was used to score all kinds of functions of 11 Qinghaosu derivatives and transferrin with different resolutions. The distances of Asp-63, Tyr-188, His-249, Arg-124 and Lys-296 with Qinghaosu were less than 0.5 nm, separately. Meanwhile, the higher is the activity of Qinghaosu derivatives the higher is the score. Our model explains that Fe2+ is more feasible to react with Qinghaosu, and not involved in other metabolism in presence of transferrin. Docking results unveil that Iron(II)-transferrin increased the cytotoxicity of Qinghaosu derivatives and provide a rational basis for further design and synthesis of novel Qinghaosu derivatives.
Antineoplastic Agents, Phytogenic ; chemical synthesis ; chemistry ; pharmacology ; Artemisinins ; chemical synthesis ; chemistry ; pharmacology ; Catalytic Domain ; Drug Discovery ; Models, Chemical ; Molecular Structure ; Protein Binding ; Transferrin ; chemistry

AIMTo study the binding of sibutramine hydrochloride (SH) and bovine serum albumin (BSA) in physiological condition by spectroscopic method.

METHODSThe quenching mechanism of the fluorescence of bovine serum albumin by sibutramine hydrochloride was studied with the fluorescence and the absorption spectroscopy. The binding constants K and the number of binding sites were determined at different temperatures according to Scatchard equation and the main binding force was discussed by thermodynamic equations. The effect of the drug on bovine serum albumin conformation was also studied by using synchronous fluorescence spectroscopy.

RESULTSThe quenching mechanism of sibutramine hydrochloride to bovine serum albumin was static quenching. The binding constants K at 8 degrees C, 25 degrees C, 37 degrees C were 1.21 x 10(5), 8.31 x 10(4), 6.97 x 10(4) L x mol(-1) with one binding site, respectively. The thermodynamic parameters of the reaction were deltaH = -9.70 kJ x mol(-1), deltaS = 56.41 J x mol(-1) x K(-1).

CONCLUSIONThe binding force is electrostatic interaction. Sibutramine hydrochloride can be deposited and transported by serum protein in vivo. Sibutramine hydrochloride has nearly no effect on the serum protein conformation.

Animals ; Binding Sites ; Cattle ; Cyclobutanes ; metabolism ; Protein Binding ; Serum Albumin, Bovine ; metabolism ; Spectrometry, Fluorescence ; methods ; Spectrophotometry, Ultraviolet ; methods

AIMTo develop a rapid and feasible method based on micellar electrokinetic capillary chromatography (MECC) for the simultaneous determination of antiepileptic drugs (AEDs)--phenytoin (PHT), phenobarbital (PB), carbamazepine (CBZ), primidone (PRM) and clonazepam (CZP) in human plasma.

METHODSSeveral factors that impact the separation of AEDs with MECC were investigated, such as concentration of sodium dodecyl sulfate (SDS), buffer compositions, pH, organic modifier, internal diameter and temperature, and an optimized MECC running condition was obtained the running buffer consisted of 8 mmol x L(-1) phosphate, 3 mmol x L(-1) sodium tetraborate, and 50 mmol x L(-1) sodium dodecylsulfate (SDS) (pH 8.0), containing acetonitrile (ACN) (18%) as organic modifier. Detection at 210 nm, run at 25 kV at 30 degrees C in a untreated fused silica capillary (50/45.5 cm length, 50 microm ID).

RESULTSThe reproducibility of both migration time and relative peak area with MECC analysis were appropriate for the intra- and inter-assay coefficients. The evaluated drugs concentration intervals of PRM 1.0-40.0 microg x mL(-1), PB 1.0-60.0 microg x mL(-1), PHT 1.0-40.0 microg x mL(-1), CBZ 1.0-40.0 microg x mL(-1), CZP 0.2-8.0 microg x mL(-1) were linear with correlation coefficients higher than 0.999 1, and coefficients of the variation of the points of the calibration curve lower than 10%. The recoveries of AEDs varied from 80.0% to 100.0%, depending on the drug, with coefficients of the variation lower than 10.0%.

CONCLUSIONThe MECC technique is showed to be rapid, simple, efficient and low cost when applied to monitoring therapeutic drugs in patient treated with a combination of PHT and other AEDs such as hepatic enzyme-inducing agents.

Anticonvulsants ; blood ; Buffers ; Carbamazepine ; blood ; Chromatography, Micellar Electrokinetic Capillary ; methods ; Clonazepam ; blood ; Epilepsy ; blood ; Humans ; Hydrogen-Ion Concentration ; Phenobarbital ; blood ; Phenytoin ; blood ; Primidone ; blood ; Sensitivity and Specificity ; Sodium Dodecyl Sulfate

AIMTo develop a near-infrared diffuse reflectance analysis (NIRDRA) method for rapid noninvasive quantitative determination of ambroxol hydrochloride in half-finished product particles and non-blister-packed, blistered tablets.

METHODSAll spectra were measured with a Fourier transform spectrometer equipped with a PbS and a InGaAs detector, an external integrating sphere, a rotating sample cup, and a fibre-optic probe for reflectance measurements. All samples were scanned from 12,000 cm-1 to 4,000 cm-1, and each sample spectrum was obtained as an automatic mean of 64 scans. No spectrum pre-processing method was used, and spectral regions, 4,602-4,247, 12,000-7,498 and 6,102-5,446, 12,000-5,446 cm-1 were selected to develope mathematical models by partial least square method for half-finished product particles and non-blister-packed, blistered tablets samples, respectively.

RESULTSThe optimal rank and mean square error determined for half-finished product particles and non-blister-packed, blistered tablets samples by cross validation method all was 6 and 0.306, 0.972 and 1.492, respectively, the average recovery was 100%, 100% and 102% respectively; and the RSD was 1.17%, 1.70% and 1.78% respectively.

CONCLUSIONResults showed that the NIRDRA method was rapid, simple, noninvasive and sensitive, and it can be applied to assay the content of ambroxol hydrochloride in half-finished product particles non-blister-packed and blistered tablets.

Ambroxol ; administration & dosage ; analysis ; Expectorants ; administration & dosage ; analysis ; Quality Control ; Spectroscopy, Near-Infrared ; methods ; Tablets

To analyze the reasons of impaired vision after LASlK and explore its preventive treatment measures preliminarily.METHODS: ln this retrospective study, 175 eyes of 134 patients whose vision was decreased after LASlK were included. The constituent ratio of every reason was counted and uncorrected visual acuity ( UCVA ) between pre-treatment and post-treatment were compared by paired t-test respectively.RESULTS:The overall incidence of impaired vision after LASlK was 1. 86%. The constituent ratio of regression was 51. 43% and UCVA increased from 0. 61±0. 22 to 0. 90±0. 38 (t=8. 00, P<0. 001) after treatment. The constituent ratio of punctate corneal epithelial defect was 32. 57% and UCVA increased from 0. 60±0. 19 to 1. 20±0. 24 (t=20. 00, P<0. 001 ) after treatment. The constituent ratio of accommodative spasm was 5. 14% and UCVA increased from 0.76±0. 21 to 1. 32±0. 22 (t=8. 14, P<0. 001) after treatment. The constituent ratio of corneal flap shift and gauffer was 4% and UCVA increased from 0. 29 ± 0. 26 to 1. 24 ± 0. 28 ( t = 6. 33, P<0. 001 ) after treatment. The constituent ratio of corticosteroid - induced ocular hypertension was 4% and UCVA increased from 0. 57±0. 05 to 1. 0 ± 0. 16 ( t= 2. 53, P<0. 05 ) after treatment. The constituent ratio of fundus lesions and diffuse lamellar keratitis ( DLK) was 2. 86% and UCVA all increased by different degrees after treatment.CONCLUSlON: The reasons of impaired vision after LASlK are many and varied. These cases could recover their vision by discovery and treatment in time, and the appropriate preventive measures were essential.

OBJECTIVEDetermine the content of ursolic acid of Liuwei Dihuangwan.

METHODUsing NIR with PLS, PCA-BPANN and WT-BPANN.

RESULTThe predication recovery were 100.7%, 100.6%, 100.1%, and the RSD were 5.42%, 6.49%, 6.52% respectively.

CONCLUSIONNIR can be used in the determination of ursolic acid, which set up the foundation of on-line control of traditional Chinese medicine.

Cornus ; chemistry ; Drug Combinations ; Drugs, Chinese Herbal ; administration & dosage ; chemistry ; isolation & purification ; Plants, Medicinal ; chemistry ; Quality Control ; Spectroscopy, Near-Infrared ; Triterpenes ; analysis