AIM: To investigate the possibility of transfecting siRNA into rabbit cervical cells in transformation zone by the method of solid phase in vivo and to verify the effectivity of siRNA transfection by modifying the permeability of the cervical epithelium. METHODS: A sense strand small-interference RNA (siRNA) for human papillomavirus type 16 (21 bp) was designed and labeled with Cy3. siRNA-Lipo2000-carbomer gum was prepared. Twelve rabbits were included in the study and divided into experimental group and control group. In order to modify the permeability of cervical epithelium, hypertonic saline solution at concentration of 200 mmol/L was used to infuse the cervix in the experimental rabbits for 10 min, and normal saline was used for the control animals. The siRNA-Lipo2000-carbomer gum was applied to the surface of the rabbit cervix. Twenty-four hours later, the rabbits were sacrificed, and the cervix was isolated, cut into 2 parts, one part was for rapid frozen sectioning and the efficiency of transfection was observed under fluorescence microscope, another part was prepared by paraffin embedding and sectioning, and the form of cervical histiocytes was observed. Twelve SCID mice with SiHa cell cervical tumor, divided into experimental group and control group, were also used in the study. The mice in experimental group were treated with siRNA-Lipo2000-carbomer gum for 7 d. The control mice were treated with Lipo2000-carbomer gum only. Five days later, the mice were sacrificed and the tumor was collected, and the HPV16-DNA was measured by PCR. RESULTS: (1) Red fluorescence (Cy3) in cervical epithelium was observed in all rabbits. However, no different effect of siRNA transfection was found between the ways of modifying the cervical epithelium permeability. (2) No abnormal change such as flare, swelling and ulcer at all cervical tissue was observed, the cervical cell form was normal. (3) The titer of HPV16-DNA was decreased significantly after siRNA transfection (P<0.05). CONCLUSION: Transfection of siRNA into rabbit cervical epithelium in vivo is successful by using the method of solid phase and inhibits the processes of HPV-DNA, indicating that using RNAi is a practical way to treat HPV infection in human cercix and to decrease the incidence of cercical carcinoma.

In this study, we investigated the pharmacokinetics parameters of SPIO-shRNA dual functional molecular probe and observed the main organ distribution by MRI in vivo. Eighteen New Zealand white rabbits were randomly divided into three groups and injected intravenously with different doses of SPIO-shRNA molecular probe, respectively. The blood samples were collected to analyze the pharmacokinetic parameters by measuring the iron content at 30 minutes before and after the injection. Twenty-four Kun Ming (KM) mice were randomly divided into 4 groups: the control group was injected intravenously with physiological saline 200 µL per mouse via the tail vein, the other 3 groups were injected intravenously with different doses of SPIO-shRNA molecular probe. MRI observation was performed in 24 hours, and the liver, spleen, kidney, brain and muscle were collected for iron quantification with Prussian blue staining to determine distribution of the SPIO-shRNA molecular probe in the main organ in vivo. Our results suggest that the molecular probe blood half-life is more than 3 hours. The data of MRI suggest the probe was distributed in liver and spleen, and the MRI signal was reduced with the increase in probe's doses (P < 0.05). The results of Prussian blue staining confirmed the results of MRI. Most of the probe could escape the phagocytosis of mononuclear phagocyte system. Our data provide the pharmacokinetic and distribution of SPIO-shRNA molecular probe in organs. Meanwhile, it suggests the choice of the time and dose of probe for MR imaging of tumor in vivo.
Animals ; Half-Life ; Magnetic Resonance Imaging ; Magnetite Nanoparticles ; Mice ; Molecular Probes ; pharmacokinetics ; RNA, Small Interfering ; chemistry ; Rabbits

Cochlear Nerve ; chemistry ; pathology ; Cranial Nerve Neoplasms ; metabolism ; pathology ; Diagnosis, Differential ; Female ; Heart Neoplasms ; metabolism ; pathology ; Humans ; Immunohistochemistry ; Middle Aged ; Neoplasms, Multiple Primary ; metabolism ; pathology ; Neurilemmoma ; metabolism ; pathology ; S100 Proteins ; metabolism ; Vestibulocochlear Nerve Diseases ; metabolism ; pathology ; Vimentin ; metabolism

OBJECTIVETo develop a new technique for assessing the risk of birth defects, which are a major cause of infant mortality and disability in many parts of the world.

METHODSThe region of interest in this study was Heshun County, the county in China with the highest rate of neural tube defects (NTDs). A hybrid particle swarm optimization/ant colony optimization (PSO/ACO) algorithm was used to quantify the probability of NTDs occurring at villages with no births. The hybrid PSO/ACO algorithm is a form of artificial intelligence adapted for hierarchical classification. It is a powerful technique for modeling complex problems involving impacts of causes.

RESULTSThe algorithm was easy to apply, with the accuracy of the results being 69.5%±7.02% at the 95% confidence level.

CONCLUSIONThe proposed method is simple to apply, has acceptable fault tolerance, and greatly enhances the accuracy of calculations.

Algorithms ; Artificial Intelligence ; China ; epidemiology ; Environmental Exposure ; adverse effects ; Humans ; Infant, Newborn ; Models, Biological ; Neural Tube Defects ; epidemiology ; Risk Factors

Objective To explore the effect of problem-based 1earning (PBL) combined with PICO model in clinical nursing internship teaching.Methods A retrospective case-control study was adopted,133 undergraduate nursing students in grade 2010 were selected as theexperimental group,and 128 undergraduate nursing students in grade 2009 were selected as the control group.The students in the experimental group received a teaching method which combined PBL with PICO model,while the students in the control group received traditional teaching methods.The two groups were evaluated by Objective Structured Clinical Examination (OSCE) and questionnaire survey.Results The OSCE grade and evaluation of teaching method in the experimental group students were higher than the control group,the difference was statistically significant.Conclusions PBL combined with PICO model teaching methods are not only beneficial to cultivate and improve the clinical nursing competency and comprehensive qualities of the nursing students,but also let the students gradually learn and permeate the ideas of any decision made by clinician should be based on the scientific research results,which from the main thought of the evidence-based practice.It can lay a good foundation for going to work in the future,and is also worthy of being popularized and applied in clinical internship teaching of undergraduate nursing students.

OBJECTIVETo apply the Wright-Giemsa stain in micronucleus test and to explore the stain outcomes of Wright-Giemsa dye of various proportions and staining times.

METHODSUse Wright-Giemsa dye, Wright dye (staining time 3 min) and Giemsa dye (staining time 5 min) to stain HepG2 and then observe the staining effect. The Wright-Giemsa dye was applied under 5 different proportions (3:1-1:3) and different staining times (1, 3, 5, 10, 15 min).

RESULTSAfter stained for 3-5 min with the proportion ratio of 3:1 of Wright-Giemsa dye, the HepG2 cells showed much better staining outcomes compared with the single stain of either Wright or Giemsa.

CONCLUSIONSWright-Giemsa stain can be used in cell micronucleus test to obtain good staining outcomes.

Azure Stains ; Coloring Agents ; Hep G2 Cells ; Humans ; Micronucleus Tests ; Staining and Labeling ; methods

AIMTo build a simple and repeatable animal model of subarachnoid hemorrhage (SAH).

METHODSSAH was produced by passing a nylon thread up through the right internal carotid artery and piercing a hole in the right anterior cerebral artery. At 12 h and 24 h after SAH operation, the rats were evaluated with rotarod test and the behavior scale (5-point scale).

RESULTSThe rats were trained through rotarod test and then randomly divided into three groups, including vehicle group treated with vehicle after SAH, nimodipine treated group (i.p. 0.25 mg x kg(-1), 5 min, 6 h, 12 h after SAH) and sham group. At the point of the perforation there was usually a capping clot. There was always blood in the basal cisterns with some spread over the hemisphere. After 12 h and 24 h of SAH operation, the time of rotarod test of rats decreased significantly and the rats had serious neurological deficit. Nimodipine could alleviate the neurological deficit after 24 h of SAH.

CONCLUSIONTo present a simple and reliable model of SAH in the rats, which allows evaluating novel compounds and new drugs for treatment of SAH.

Animals ; Behavior, Animal ; drug effects ; Brain ; pathology ; Calcium Channel Blockers ; pharmacology ; Cerebrovascular Circulation ; drug effects ; Disease Models, Animal ; Male ; Nimodipine ; pharmacology ; Rats ; Rats, Wistar ; Reproducibility of Results ; Rotarod Performance Test ; Subarachnoid Hemorrhage ; etiology ; pathology ; physiopathology

This study aimed to construct the dual expression vectors of wide type or N187D mutant P2X7 receptor and intracellular domain of Notch1 (ICN1) linked by 2A peptide to coexpress them in leukemia cells so as to lay a foundation for further investigating the role of P2X7 in development of leukemia. Overlap PCR was used to construct the dual expression vectors encoding wide type or N187D mutant type P2X7 receptor and ICN1 linked by the self-cleaving 2A sequence. The results showed that stable expressing cell lines were obtained by retroviral infection followed by cell sorting after DNA sequence analysis. RT-PCR, Western blot, intracellular free calcium concentration analysis were used to verify the functionally successful construction of K562 cell line expressing P2X7 receptor alone or with ICN1. DNA sequence analysis revealed that all construction were right. The infection efficiency of packaged constructed virus ranged from 40% to 70% for K562 cells. Stable infected cell line was obtained by cell sorting. RT-PCR analysis revealed that P2X7 receptor and/or ICN1 could be detected at high level in their stable infected cell lines, respectively. Western blot analysis also showed that P2X7 receptor was highly expressed in cell line infected by virus with P2X7 receptor. Sustained increase in intracellular free calcium concentration ([Ca(2+)]i) could be observed in K562 cells overexpressing either type of P2X7 receptor upon stimulation with BzATP. It is concluded that the wide type or N187D mutant P2X7 receptor and ICN1 are simultaneously and functionally over-express in leukemia cells, which lay a foundation for further studying the role of P2X7 receptor in the development of leukemia.
Gene Expression ; Genetic Vectors ; Humans ; K562 Cells ; RNA, Messenger ; genetics ; Receptor, Notch1 ; genetics ; Receptors, Purinergic P2X7 ; genetics ; Retroviridae ; genetics

OBJECTIVETo establish a new method for simultaneous determination of shanzhiside methyl ester, chlorogenic acid, 8-O-acetyl shanzhiside methylester, forsythiaside B, rutin, acteoside and galuteolin in Lamiophlomis rotata.

METHODSeparation was performed on a Welchrom-C18 chromatographic column with acetonitrile-0.1% orthophosphoric acid as mobile phasewith gradient elution. The flow rate was 1.0 mL x min(-1). The column temperature was 30 degrees C, and the detection wavelength was set at 238 nm, 330 nm and 350 nm.

RESULTThe seven compounds were well separated with good linear correlations. The mean recoveries of seven compounds were 96.47%-102.2% (RSD 0.70%-2.2%).

CONCLUSIONThere were good correlations among the seven compounds in the samples of aerial parts. The mean sum of shanzhiside methyl ester and 8-O-acetyl shanzhiside methylester in samples of aerial parts is 1.44%. The aerial parts have more kinds of composition and with higher content than that of underground parts in L. rotata, which was reasonable for the resonable use of the aerial part as medicinal part. The method was simple, repeatable and stable, which could be used for identification and quality evaluation of L. rotata.

Chlorogenic Acid ; chemistry ; Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; chemistry ; Glucosides ; chemistry ; Glycosides ; chemistry ; Lamiaceae ; chemistry ; Methyl Ethers ; chemistry ; Phenols ; chemistry ; Plant Components, Aerial ; chemistry ; Plants, Medicinal ; chemistry ; Pyrans ; chemistry ; Rutin ; chemistry

OBJECTIVETo explore the feasibility of autologous burn eschar as the coverage of microskin grafting in the repair of excised severe burn wounds.

METHODSTwelve severe burn patients underwent massive escharectomy during 3 to 7 post burn days (PBD), and autologous eschar instead of alloskin was employed as the coverage of microskin. The integrity of grafted eschar and survival of microskin grafts were observed at 1 to 6 weeks after operation. The wound healing rate in grafted area was determined at 6 post operation week (POW), and the wound healing time was recorded.

RESULTSThe autologous eschar in the grafted area were integral and attached tightly to the wound during the 1 to 2 POW and began to dry and detach from the burn wound with the microskin underneath growing and fusing at 3-4 POW. This process went on until the eschar was completely detached from the burn wound and the microskin fused in large areas, with some granulation wounds left at 5 and 6 POW. The wound healing rate at the 6 POW was (87 +/- 4)%. The average wound healing time of burn patients in this group was (56 +/- 8) days.

CONCLUSIONAutologous eschar could be used as a substitute for the alloskin for microskin grafting in excised burn wounds.

Adolescent ; Adult ; Burns ; surgery ; Feasibility Studies ; Female ; Graft Survival ; Humans ; Male ; Reconstructive Surgical Procedures ; Skin Transplantation ; methods ; Surgical Flaps ; Transplantation, Homologous ; Wound Healing