Animals ; Benchmarking ; methods ; statistics & numerical data ; Dose-Response Relationship, Drug ; Humans ; Models, Statistical ; No-Observed-Adverse-Effect Level ; Risk Assessment ; methods ; statistics & numerical data ; Toxicology ; methods

Adult ; Arsenic ; pharmacology ; Chromosome Aberrations ; chemically induced ; Dose-Response Relationship, Drug ; Female ; Humans ; Lymphocytes ; drug effects ; metabolism ; Male ; Middle Aged

To determine the optimum process conditions for dry granulating technique of Qibai Pingfei granule, granule excipient type, rolling wheel speed and pressure and feeding speed were studied. Taking shaping rate at a time, moisture absorption and dissolubility as index, the type and amount of granule excipient were determined. In addition, taking shaping rate at a time as index, parameters of rolling wheel speed and pressure and feeding speed were researched through single factor test and response surface methodology. The optimum parameters were as follows: lactose as excipient, dry extract powder to excipient at 1:2, rolling wheel speed and pressure at 10.9 Hz and 6.4 MPa and feeding speed at 7.2 Hz. After validation of three batches pilot-scale production, the optimum processing parameters for dry granulating technique of Qibai Pingfei granule is reasonable and feasible, which can provide reliable basis for production.
Drugs, Chinese Herbal ; Powders ; Technology, Pharmaceutical ; methods

Sanjie Zhentong capsules were scanned by using a near infrared spectra probe with different drug mass fraction and the spectral information of capsule shells and contents in it were obtained. Then partial least squares (PLS) models were developed for the prediction of mass fraction of Fritillariae Thunbergii Bulbus and Resine draconis in Sanjie Zhentong capsules. The correlation coefficient (r9c)) and root mean standard error( RMSEC) of 0.949 5, 0.958 2 and 4.742 4, 4.135 7. The models obtained correlation coefficient (r(v)) of 0.919 2, 0.936 7 and root mean square error (RMSECV) of 6.158 9, 5.037 3 respectively in the training set. The paired T test analysis of statistics showed that there were no significant difference between predictive values and measure values. The established models reflected a strong prediction performance and can meet the needs of the production.
Capsules ; chemistry ; Drugs, Chinese Herbal ; chemistry ; Least-Squares Analysis ; Spectroscopy, Near-Infrared ; methods

Chronic osteomyelitis is one of the most common disorder in clinic. In recent years due to diabetes, peripheral vascular disease and trauma induced disease increased, the prevalence rate increased. With the development of magnetic resonance imaging and CT imaging technology, it greatly improved the accuracy of clinical diagnosis of chronic osteomyclitis and ability to describe the infection characteristics, and provide a reliable basis for clinical treatment. The current research on chronic osteomyelitis mainly concentrated on the aspects of imaging applications and ways of using antibiotic optimization control inflammation, defect restoration and reconstruction of blood supply and treatment. But the best time to the antibiotic therapy and the use of program is still uncertain, for after debridement, bone grafting time and defect repair function of fast recovery still need further research.
Anti-Bacterial Agents ; therapeutic use ; Chronic Disease ; Humans ; Osteomyelitis ; diagnosis ; therapy

OBJECTIVETo observe effects of serum Bushen Huoxue prescription(Chinese characters) on classic Wnt/β-catenin signaling pathways of osteoblasts, and explore mechanism of Bushen Huoxue prescription (Chinese characters) for preventing osteoporosis.

METHODSTwenty health female rats were randomly divided into two groups, including Bushen Huoxue (Chinese characters) group and saline group,10 in each group. Bushen Huoxue (Chinese characters) group and Saline group were gavaged Bushen Huoxue and saline every day for 1 week. Bushenhuoxue containing serum and saline containing serum were got according to methods of serum preparation of drug-containing. The osteoblasts was cultured with neonatal rat skull according to Enzyme Consumer Law, and was identified by Wright-Giemsa staining (R-J) and alkaline phosphatase staining (ALP). The third generation of osteoblasts was divided into three groups, including saline group, normal group,Bushen Huoxue (Chinese characters) group. Each group were added to 15% appropriate medium. ALP activity of osteoblasts and osteoblasts proliferation rate were tested, mineralized nodules were observed, the expression of β-catenin, Runx2, Osx mRNA of osteoblasts were tested by RT-PCR.

RESULTSThere was blue granules in cytoplasm, cell nucleus was flint with 1 to 3 nucleoli showed by R-J staining, morphology of osteoblasts were cultured. ALP staining showed cytoplasm with purple granules, the results showed that the cultured cell was osteoblasts. The content of ALP in Bushen Huoxue (Chinese characters)group was (6.272±0.131) ,appreciation rate was (0.81? 0.172), and could significantly improve differentiation and proliferation activity of osteoblasts compared with Saline group (P< 0.01). There were four different size orange nodules, the Maximun nodule was 1.0 x 1.0 cm in Bushen Huoxue (Chinese characters) group after Alizarin red staining, the results showed Bushen Huoxue (Chinese characters)group could obviously improve mineralization of osteoblasts. The expression of mRNA of β-catenin, Runx2 and Osx in Bushen Huoxue (Chinese characters)group were (1.782±0.944), (1.935±0.994) and (1.610±0.811) by RT-PCR,it was significantly increased compared with saline group (P<0.01), but there was no difference between Bushen Huoxue (Chinese characters)group and normal group (P>0.05).

CONCLUSIONBushen Huoxue (Chinese characters)group could obviously promote differentiation, proliferation and mineralization of osteoblasts through activation of Wnt, β-catenin signaling pathway. It suggested that the mechanism of action of Bushen Huoxue (O'f f Il.t)particle clould prevent osteoporosis through the activation of Wnt, β-catenin signaling pathway.

Animals ; Cell Differentiation ; drug effects ; Cell Proliferation ; drug effects ; Drugs, Chinese Herbal ; pharmacology ; Female ; Humans ; Osteoblasts ; cytology ; drug effects ; metabolism ; Osteoporosis ; drug therapy ; genetics ; metabolism ; physiopathology ; Rats ; Rats, Sprague-Dawley ; Wnt Proteins ; genetics ; metabolism ; Wnt Signaling Pathway ; drug effects ; beta Catenin ; genetics ; metabolism

Objective To investigate the effects of baicalein and acitretin on the apoptosis in a human cutaneous squamous cell carcinoma cell line, SCL-12. Methods Cultured SCL-12 cells were treated with different concentrations of baicalein (3.125, 6.25, 12.5 μmol/L) and acitretin (2.5, 5.0, 10.0 μ mol/L), alone or in combination, for 48 hours. Subsequently, cell proliferation was detected by MTT assay, and cell apoptosis by ELISA as well as annexin V-FITC and propidium iodide double staining. Real-time quantitative RT-PCR was used to detect the expression of Fas mRNA in SCL-12 cells. Results The cell proliferation of SCL-12 cells was inhibited by baicalein and acitretin alone or in combination. The combination of baicalein and acitretin at the three tested concentrations, except for that of baicalein at 3.125 μmol/L and acitretin at 2.5 μmol/L, more strongly inhibited the proliferation of SCL-12 cells compared with baicalein or acitretin alone, and the inhibitory effect was in a dose-dependent manner. The early apoptosis rate was 9.39% ± 1.52%, 20.86% ± 2.16%,36.85% ± 3.26% in SCL-12 cells treated with baicalein of 3.125 μmol/L, acitretin of 5.0 μmol/L alone and their combination, respectively, significantly higher than that in untreated cells (4.39% ± 0.64%, all P <0.05); the induction of apoptosis in SCL-12 cells by the combination of baicalein and acitretin was stronger than that by baicalein or acitretin alone (F = 138.44, P < 0.05). Baicalein and acitretin alone or in combination significantly increased the mRNA expression of Fas in SCL-12 cells, and the effect of their combination was stronger than that of baicalein or acitretin alone. Conclusions Baicalein and aeitretin could inhibit the growth of and induce the apoptosis in SCL-12 cells, and the effect is enhanced by the combination of baicalein and acitretin, which may be associated with the upregulation of Fas expression in SCL-12 cells.

Objective To develop a method of real-time quantitative polymerase chain reaction (RQ-PCR) for detection of translocation ETS leukemia-acute myeloid leukemia 1(TEL-AML1) fusion gene in children with acute lymphoblast leukemia(ALL),and explore its clinical application in minimal residual disease (MRD) monitoring.Methods By reverse transcription and RQ-PCR,a quantifying of TEL-AML1 fusion gene was developed,and the expression levels of TEL-AML1 were detected in bone marrow (BM) samples obtained from 24 children with ALL at diagnosis and at the end of induction of remission,as well as at a series of follow-up. Moreover,the results of MRD detection by RQ-PCR were compared with that of detection by routine morphological examine of BM and cells differentiation mark analysis by flow cytometer(FCM),to evaluate the sensitivity of RQ-PCR in MRD monitoring. Results A method of RQ-PCR targeted at TEL-AML1 fusion gene was established. In 11 BM samples,which collected from TEL-AML1 positive children at the end of induction therapy and all of them achieved completely remission (CR) by routine morphological examine,5 samples were found to be MRD positive by RQ-PCR,positive ratio was 45%. There were 15 BM samples collected in maintenance therapy period,and all these samples were CR by routine morphological examine. However,by RQ-PCR,3 out of 15 samples were found to be MRD positive during maintenance therapy period. After intense and maintenance therapy,the MRD levels of the 3 children were declined to negative. In a recrudescent child,the expression of TEL-AML1 fusion gene was rose by a magnitude of 103 copies before relapse,and after induction therapy once again the patient was completely relaxed.Conclusions RQ-PCR targeted at TEL-AML1 has a higher sensitivity than conventional morphologic way and FCM. RQ-PCR can be used in the quantitative detection of MRD,and provide gist for early prognosticating a relapse and instructing clinical therapy.

AIM: In order to explore the pathogenesis of ulcerative colitis (UC), an experimental colitis in mouse was induced by the hapten dinitrochlorobenzene (DNCB), and the activity of leukocyte migration inhibitory factor (LMIF) was measured at the same time. METHODS: 67 BALB/c mice were randomly divided into control (60% ethanol) and DNCB groups. After they were sensitized by smearing 3.3% DNCB on the abdominal skin, they were challenged with DNCB at concentration of 0.1%, 0.2% and 0.4% respectively by instillation once a day. The weight, stool viscosity and hematochezia were observed and accumulated as disease active index (DAI) score. The pathological changes in colon tissue were judged macropathologically and by means of microscope. LMIF activity was determined by the absorbance (A) of migrated leukocytes. RESULTS: Compared to control group, the increases in DAI accumulate score, pathologic score, and LMIF activity in DNCB groups were observed. CONCLUSION: Mouse colitis was induced by DNCB, which was accompanied by an increase in LMIF activity. [

Natural products are important sources of drug discovery. However, the traditional methods of extraction and isolation, as well as chemical synthesis for obtaining natural products are associated with issues such as operational complexity, high costs, low efficiency, and environmental pollution. Constructing microbial cell factories through synthetic biology methods to produce medicinal natural products has the advantages of high efficiency, low cost, and environmental protection. Nevertheless, the scope and yield improvement of the products are limited by the limitations of enzymes in microbial cell factories. Protein engineering is considered one of the most effective approaches to overcome these limitations. This article introduces commonly used methods of protein engineering technology and summarizes its specific applications in improving enzyme performance, modifying the enzymatic environment, and promoting the development of synthetic biology tools in the field of pharmaceutical natural product synthesis. Furthermore, it analyzes the current bottlenecks and challenges in protein engineering and looks forward to its future application prospects, offering insights for the development and practical use of protein engineering technology.