Objective A quantitative study of the DNA skeletal muscle cell nuclei during 0～96h postmortem was performed.Method By the histochemical and image analysis.Result The color and the areas of the nuclei stained with the Feulgen staining method became fainter and smaller gradually at 12 hpm. There was a linear relationship between the degradation rate of the nuclear DNA and the length of the PMI.Conclusion The quantitative image analysis of DNA has a wide prospect of application, but the method has been studied further in aspects of the precise and standadization of the method as well as the effects exerted on the results from the study materials.

Objective: To investigate the effects of HPV16E6-ribozyme on telomerase activity in cervical carcinoma cell line CaSKi and the related mechanisms. Methods: Anti-HPV16E6-ribozyme and blank eucaryotic plasmids were transfected into CaSKi cells via lipofectin, and the resultant cells were named as CaSKi-R and CaSKi-P, respectively. The expression of ribozyme in transfected cells was observed by RNA dot blotting. The expression of E6 mRNA and protein in the 3 kinds of cells were detected by Northern blotting and Western blotting, respectively. Telomerase activity was determined by TRAP-Elisa method; the expression of P53, c-myc, hTERT and hRT mRNA were examined by RT-PCR.Results: RNA dot blotting showed that anti-HPV16E6-ribozyme was stably expressed in transfected CaSKi-R cells. Western blotting showed that the expression of E6 mRNA and protein in CaSKi-R cells was obviously lower than that in CaSKi and CaSKi-P cells. The telomerase activities in CaSKi,CaSKi-P and CaSKi-R cells were (0.89?0.14), (0.90?0.11) and(0.36?0.06),respectively. The inhibitory rate of telomerase activity in CaSKi-R cells was 59.55%, which was significantly lower than those in CaSKi and CaSKi-P cells (P

Objective: To investigate the characteristics of the cultured cervical cancer cell line transfected with anti HPV16E6 ribozyme, and to investigate the possibility and practicality of ribozyme in treatment of cervical cancer. Methods: The anti HPV16E6 ribozyme and empty eucaryotic expressing plasmids were transfected by lipofectin transfection into CaSKi cell, which named as CaSKi R, CaSKi P respectively. The morphology and the soft agar forming ability were studied. The expression of E6, PCNA and C erbB 2 genes was studied through Flow Cytometry. The tumorgenicity of each cell was detected by injecting cells into the nude mice skin. Three groups of nude mice were injected by CaSKi, CaSKi R and CaSKi P cell separately. Another group of mice was injected by CaSKi cell on right side and CaSKi R cell on left side. Results: There is no distinct difference of the morphology and growth rate between CaSKi and CaSKi P, but the growth rate of CaSKi R decreased. The soft agar forming rate of CaSKi P was similar with that of CaSKi cells, while that of CaSKi R was found decreased. The result of flow cytometric analysis showed that anti HPV16E6 ribozyme could reduce the expression of E6, PCNA and C erbB 2 genes on CaSKi R cells, while this phenomenon was not found on the CaSKi P cells. The tumorgenicity of CaSKi R in nude mice was decreased compared with CaSKi and CaSKi P cells. Conclusion: Anti HPVE6 rivozyme could partly reverse the malignant phenotypes of CaSKi cells. The reason may be the decrease of E6 gene expression, and the succeeding decrease of the PCNA and C erbB 2 genes′ expression.

Child ; Humans ; Male ; Paralyses, Familial Periodic ; diagnosis ; therapy

OBJECTIVE: To analyze and summarize clinical characteristic, treatment scheme and survival rate of primary melanom in nasal cavity and sinus. METHOD: We retrospectively analyzed the 9 patients with primary melanom in nasal cavity and sinus who in data proceed were treated and reviewed the related literature. RESULT: Among the 9 patients, the clinical main symptoms are rhinostegnosis of lateral nasal and intermittent nasal bleeding. Pathologic examination is mainly characterized by tumor cells abnormity and cytoplasm containing pigment or without pigment, and main diagnosis basis is some or all of the positive for HMB45, S-100, melan-A. The survival rate are 88.9% (8/9) of 1-year, 66.7% (6/9) of 3-year and 33.3% (3/9) of 5-year. CONCLUSION The incidence of primary melanom in nasal cavity and sinusis is not frequent in clinic and confirmed by immunohistochemical. The extensive radical excision of focus and combine adjuvant radiotherapy postoperative may improve the survival rate of patients.
Aged ; Aged, 80 and over ; Female ; Follow-Up Studies ; Humans ; Male ; Melanoma ; diagnosis ; surgery ; Middle Aged ; Nasal Cavity ; Nose Neoplasms ; diagnosis ; surgery ; Paranasal Sinus Neoplasms ; diagnosis ; surgery ; Retrospective Studies

The study is aimed to explore a rapid method to extract DNA from fried Chinese medicinal products. The alkaline lysis buffer was made of sodium hydroxide, 1% PVP and 1% TritonX-100 and Tris-HCl solution was neutralized, through heat cracking and neutralization two step to extract DNA from processed and prepared products of traditional Chinese medicine. Then universal primes were used to amplify PCR products for fired Chinese medicinal materials. The results indicated the optimized alkaline lysis method for extracting DNA is quick and easy. Extracting of the different processed Sophora japonica of DNA concentration was (420.61 ± 123.91) g x L(-1). Using 5% Chelex-100 resin purification can improve the DNA concentration. Our results showed that the optimized alkaline lysis method is suitable for Chinese medicinal materials for quickly DNA extraction.
Alkalies ; chemistry ; Chemical Fractionation ; methods ; DNA, Plant ; genetics ; isolation & purification ; Hydrolysis ; Plants, Medicinal ; chemistry ; classification ; genetics ; Polymerase Chain Reaction ; Sophora ; chemistry ; classification ; genetics

Objective To investigate the effects of electrical stimulation on the expression of glial fibrillary acidic protein (GFAP) andinterleukin-1 alpha (IL-1α) in adult rats with spinal cord injury. Methods 72 adult SD rats were randomly divided into damage group (n=24), electrical stimulation group (n=24) and normal group (n=24). The spinal cord incomplete injury model on T9 was made with Allen'smethod in the former 2 groups. The rats in electrical stimulation group accepted electrical stimulation for 7 d. All the rats were evaluatedwith the Basso, Beattie & Bresnahan locomotor rating scale (BBB scale), and the expression of GFAP and IL-1α were determined with immunohistochemistry.Results The BBB scores in both the damage group and electrical stimulation group were significantly less than that inthe normal group (P<0.05), and it was more in the electrical stimulation group than in the damage group 5 and 7 d after injury. The expressionsof the GFAP significantly increased after injury to the peak on 5th day, while it was less in the electrical stimulation group than in thedamage group 5 and 7 d after injury (P<0.05). The expressions of the IL-1α increased continually after injury, while it was less in the electricalstimulation group than in the damage group 5 and 7 d after injury (P<0.05). Conclusion Electrical stimulation can inhibit the expressionof GFAP and IL-1α, that reduce inflammation and glial scar formation.

Adult ; Autonomic Nervous System Diseases ; etiology ; physiopathology ; Biopsy ; adverse effects ; Hepatitis B, Chronic ; pathology ; physiopathology ; Humans ; Liver ; pathology ; physiopathology ; Male ; Vagus Nerve ; physiopathology ; Vasomotor System ; physiopathology

To build the Dendrobium nobile -T2DM network, and elucidate the molecular mechanism of D. nobile to type 2 diabetes (T2DM). Collect the chemical composition of D. nobile and the targets on T2DM by retrieving database and documents, build the network of D. nobile to T2DM using the entity grammar systems inference rules. The molecular mechanism of D. nobile to T2DM includes: (1) regulating lipid metabolism by lowering triglyceride; (2) reducing insulin resistance; (3) protecting islet cells; (4) promoting the glucose-dependent insulin tropic peptide (GIP) secretion; (5) inhibiting calcium channel. Under the guidance of network pharmacology, through entity grammar systems inference rules we elucidate the molecular mechanism of D. nobile to T2DM, and provide the basis for the further development of health care products based on D. nobile.
Animals ; Calcium Channels ; genetics ; metabolism ; Databases, Factual ; Dendrobium ; chemistry ; Diabetes Mellitus, Type 2 ; drug therapy ; genetics ; metabolism ; Drugs, Chinese Herbal ; administration & dosage ; chemistry ; Gene Regulatory Networks ; drug effects ; Humans ; Hypoglycemic Agents ; administration & dosage ; chemistry ; Insulin Resistance ; Islets of Langerhans ; metabolism ; Triglycerides ; metabolism

In order to solve the adhesion and the softening problems of traditional Chinese medicine extract during spray drying, a new method of adding dehumidified air into spray drying process was proposed, and the storage stability conditions of extract powder could be predicted. Kouyanqing extract was taken as model drug to investigate on the wet air (RH = 70%) and dry air conditions of spray drying. Under the dry air condition, the influence of the spray drying result with different air compression ratio and the spray-dried powder properties (extract powder recovery rate, adhesion percentage, water content, angle of repose, compression ratio, particle size and distribution) with 100, 110, 120, 130, 140 °C inlet temperature were studied. The hygroscopic investigation and Tg value with different moisture content of ideal powder were determined. The water activity-equilibrium moisture content (aw-EMC) and the equilibrium moisture content-Tg (EMC-Tg) relationships were fitted by GAB equation and Gordon-Taylor model respectively, and the state diagram of kouyanqing powder was obtained to guide the rational storage conditions. The study found that in the condition of dry air, the extract powder water content decreased with the increase of air compression ratio and the spray drying effect with air compression ratio of 100% was the best performance; in the condition of wet air, the extract powder with high water content and low yield, and the value were 4.26% and 16.73 °C, while, in the dry air condition the values were 2.43% and 24.86 °C with the same other instru- ment parameters. From the analysis of kouyanqing powder state diagram, in order to keep the stability, the critical water content of 3.42% and the critical water content of 0.188. As the water decreased Tg value of extract powder is the major problem of causing adhesion and softening during spray drying, it is meaningful to aid dehumidified air during the process.
Drug Stability ; Drugs, Chinese Herbal ; chemistry ; Humidity ; Medicine, Chinese Traditional ; Plant Extracts ; chemistry ; Powders ; Temperature