AIM:Neural stem cells can be induced to differentiate into various types of neural cells such as neurons and neuroglia cells,but the technique of depuration and cultivation does not consummate.This article determines the optimal culture technique of neural stem cells by different culture concentrations and passage methods. METHODS:Experiments were conducted from May to December 2006 at Laboratory of Transplantation Immunity of Sichuan University.①Clean pregnant female rats(embryonic age range from 12-16 days)and the disposition of animal met ethical standard.②The cerebral cortex of rat embryos were collected,and digested with trypsin and ethylenediamine tetraacetic acid mixture to obtain signal cell suspension.They were cultured in serum-free medium (DMEM/F12 medium containing B27,basic fibroblast growth factor and epidermal growth factor).The 3~(rd)passage cells were collected,and incubated at 1?10~7 L~(-1),1?10~8 L~(-1),1?10~9 L~(-1),1?10~(10)L~(-1),respectively.In addition,neural stem cells were collected 7-10 days after primary culture to harvest formative cell masses.Mechanical blow refers to soft blowing with haustorial tube from thick to thin after centrifugation,or sterile syringe with No.5 pinhead blowing cells when the blow was 5 times.Bubble production was avoided during the operation.Trypsin aspiration combined with mechanical blow refers to trypsin was added after centrifugation,at 37℃for 10 minutes,the neural stem cells were lightly blown with haustorial tube polished with flame or blown with sterile syringe with No.5 pinhead,and then fetal bovine serum was added to stop digestion.③The growth characteristic of the 3~(rd)passage cells at different culture concentration was observed and proliferation was measured at days 1,3,5 and 7 by MTT assay.The neural clone spheres of subcultured was counted to determine the optimal passage way.Immunofluorescence was carried out to detect nestin(special marker to neural stem cells),BrdU,neurone specific enolase,glial fibrillary acidic protein. RESULTS:①Growth characteristics and identification of rat embryonic neural stem cells in vitro:The dissociated neural stem cells from the cerebral cortex of rat embryos were continuously harvested and purified by suspension cultures to get the daughter cell clone.Nestin positive cells could be found in the neurospheres and after attachment they could differentiate into neurone specific enolase and glial fibrillary acidic protein positive cells,and immunofluorescence showed a great of BrdU-positive cells.②The effect of different incubated number of neural stem cells on proliferation:When the neural stem cells planted at the concentration of 1?10~9 L~(-1),the growth rate of the cells was the highest of all concentrations.The number of clone spheres exceeded others at concentration 1?10~7 L~(-1),1?10~8 L~(-1)and 1?10~10 L~(-1)(P

Objective To observe the clinical characteristics and surgical methods of strabismus following external approach for retinal detachment,and explore the common factors related to deviation angles of strabismus.Methods Twenty-five cases of secondary strabismus following external approach for retinal detachment underwent strabismus surgery were recruited,and the follow-up time was from 5 months to 41 months.Ophthalmic examinations were performed before operation.Clinical characteristics of these cases were observed by intraoperative exploration,including preoperative and postoperative eye position and ocular movement (at 1 week and further followup).The preoperative and postoperative deviation angles of strabismus were evaluated according to the times of retinal detachment operation,postoperative BCVA and the surgery procedure of hang-back recession.Results Before strabismus operation,all cases underwent abnormal eye-movements,8 cases suffered from diplopia,and scar adhesions were observed in all involved ocular muscles during operation.Seven cases underwent weakening procedure of extraocular muscle,18 cases underwent weakening procedure plus strengthening procedure.Ten cases underwent hang-back recession among them.At 1 week and the final follow-up,the eyeball in21 cases and 15 cases were in position,4 cases and 9 cases were in undercorrection,5 cases and 7 cases were in limitation,and 2 cases and 1 case were in diplopia.The more operations were performed in retinal detachment,the more scar adhesions were presented,and statistical significance existed with the mean deviation angles before and after operation(in a week) between cases that had one-time and over-one-time of retinal detachment surgery (P =0.01,0.00).No difference of mean deviation angles was found at one week after operation and at follow-ups between the cases with and without hang-back recession (P =0.26,0.63).Conclusion Abnormal eyemovements,diplopia and scar adhesion are the most common clinical characteristics of strabismus following retinal detachment surgery.Times of retinal detachment operation and postoperative BCVA can influence the deviation angles of strabismus,and hang-back recession seems the safe and effective method of strabismus surgery.

Objective To research the absorption mechanism of paeoniflorin across Caco-2 monolayer model. Methods Depending on the Caco-2 cell monolayers drug transport model to study the double transport mechanism of paeoniflorin. To explore the effect of paeoniflorin absorption by time and drug concentration, the drug concentration was determined through the use of HPLC instrument and the Papp was calcalatad. Results In the Caco-2 monolayer model, the transport of paeoniflorin form Apical (AP) to Basolateral (BL) is similar to the transport form BL to AP. Conclusion The main mechanism of the paeoniflorin intestinal absorption in the Caco-2 monolayer model is passive transference.

Objective To investigate the role of Rho/Rho kinase -associated cell migration of hepatic carcinoma cell line and inhibition of tumor cell invasion by Rho kinase inhibitor Y-27632. Methods Western blot was used to estimate the expression of Rho protein in the cells. After treatment of SMMC7721 cell with Y-27632, cell biological behaviors such as colony-forming efficiency, adhesiveness, cell motility, in vitro invasiveness, metastatic potential were observed. Results The ability of Y-27632 treated SMMC7721 cells to invade the reconstituted basement membrane decreased significantly ( P

Objective To evaluate the acute side effects and efficacy of three dimensional conformal radiotherapy(3DCRT) combined with chemotherapy(CT) for locally advanced non small cell lung cancer(LANSCLC). Methods From Sept. 2000 to Dec.2001,24 patients with LANSCLC were treated with three dimensional conformal radiotherapy combined with chemotherapy navelbine+cisplatin NP. Results The overall response rate(CR+PR)was 91.7% for the primary tumor, and 100% for metastatic mediastinal lymph nodes.The leucopenia rate was 95.8%(54.2% grades 3,4). Acute radiation esophagitis occurred in 54.2% of patients(Grades 1,2). Acute radiation pneumonitis was observed in 12.5% of patients (Grades1,2). Conclusions 3DCRT combined with CT is well tolerated in most patients with LANSCLC. Its early responses are milder than the conventional radiotherapy combined with chemotherapy . Remote results await further follow up.

OBJECTIVE:To probe into the cost control in PIVAS of our hospital in order to provide reference for the effective cost control.METHODS:The status quo of cost control of PIVAS was analyzed to provide corresponding countermeasures.RESULTS & CONCLUSIONS:The effective cost control can be achieved through improving cost accounting,controlling cost and reducing running cost and labour cost to promote the healthy development of PIVAS.

In this study, a detailed L3-L5, three dimensional nonlinear finite element model of a normal aged 32 man was established with the aid of two softwares, CATIAV5 and MSC. Marc, which can derive a three-dimensional finite element model from CT images. We loaded the model with 10 N * m net moment and computed the average rigidity of functional spinal unit. The model was validated in contrast to the former biomechanical studies of cadaveric spine. The model will be used in further clinical biomechanical studies.
Adult ; Biomechanical Phenomena ; Finite Element Analysis ; Humans ; Imaging, Three-Dimensional ; methods ; Lumbar Vertebrae ; diagnostic imaging ; physiology ; Male ; Models, Biological ; Nonlinear Dynamics ; Radiography ; Stress, Mechanical

Objective To investigate antibiotics resistant characteristics and carbapenemases genotype of Acinetobacter baumannii in Intensive Care Unit (ICU),so as to provide theoretical basis for clinical prevention and treatment.Methods Retrospective study was made on 90 non-duplicated clinical isolates of Acinetobacter baumannii,which were collected From January 2013 to January 2014 in three tertiary hospitals of Qingdao.All strains were identified by VITEK2 automated microbiology analyzer;K-B method was used to do drug susceptibility test;polymerase chain reaction (PCR) was used to amplify the OXA-23,OXA-24,OXA-51,OXA-58,KPC-2,VIM,IMP genes,and the positive products of genes were sequenced;the chi-square test was used to compare the difference of the resistance rates.Results The detection rate of multi-drug resistant A.baumannii (MDRAB)and Pan-drug resistant A.baumannii (PDRAB)was 61.11% (55/90) and 17.78% (16/90).In the 32 strains of imipenem-resistant Acinetobacter baumannii,the resistant rates to Cefoperazone/sulbactam,Polymyxin B was lower,while the resistant rates to other drugs tested were more than 85%.The difference of the resistance rates to 9 drugs between imipenem resistant group and Imipenem sensitive group were statistically significant (P≤0.05).PCR result showed: 32 strains detected OXA-51 gene,28 strains detected OXA-23 gene,and 3 strains detected VIM gene,the detection rates of which were 100%,87.50% and 9.38% respectively.All strains were not detected OXA-24,OXA-58,KPC-2 and IMP genes.The sequenced results were absolutely homology with the corresponding genes in genbank.Conclusions The resistance of A.baumannii in ICU is serious in this region,especially imipenem-resistant A.baumannii,which were nearly no-sensitive to most of the drugs commonly used in clinical.The gene existence of carbapenemase and carbapenemase producing is one of the main resistance mechanism of Acinetobacter baumannii to carbapenem antibiotics.OXA-23 was the major genotypes in this region.

Objective To design a novel electronic device for measuring the pressure in the cuff of the artificial airway; and to study the advantage of this device on continuous and intermittent cuff pressure monitoring. Methods ① a portable electronic device for cuff pressure measurement was invented, which could turn pressure signal into electrical signal through a pressure transducer. Meantime, it was possible to avoid pressure leak from the joint and the inside of the apparatus by modified Luer taper and sophisticated design. If the cuff pressure was out of the normal range, the apparatus could release a sound and light alarm. ② Six traditional mechanical manometers were used to determine the cuff pressure in 6 tracheal tubes. The cuff pressure was maintain at 30 cmH2O (1 cmH2O =0.098 kPa) by the manometer first, and repeated every 30 seconds for 4 times. ③ Study of continuous cuff pressure monitoring: We used a random number generator to randomize 6 tracheal tubes, 6 mechanical manometers and 6 our products by number 1-6, which has the same number of a group. Every group was further randomized into two balanced groups, one group used the mechanical manometer first, and the other used our product first. The baseline pressure was 30 cmH2O, measurement was performed every 4 hours for 6 times. Results When traditional mechanical manometer was used for cuff pressure monitoring, cuff pressure was decreased by an average of 2.9 cmH2O for each measurement (F = 728.2, P = 0.000). In study of continually monitoring, at each monitoring point, the pressure measured by electronic manometer was higher than the mechanical manometer. All the pressures measured by mechanical manometer were dropped below 20 cmH2O at 8th hour, and there was no pressure decrease below 20 cmH2O measured by electronic manometer in 24 hours by contrast. In study of intermittent monitoring, the same result was found. The pressure was dropped significantly with time when measured by mechanical manometer (F = 61.795, P = 0.000), the drops below 20 cmH2O began at 8th hour; but when measured by electronic manometer, all the value stayed unchanged around the baseline in 24 hours (F = 0.511, P = 0.796). Conclusions Compared with traditional mechanical manometer, cuff pressures monitored by our novel electronic manometer were steadier in both continuous and intermittent monitoring. The device is compact and convenient, and can provide a good solution for continuously monitor of the tracheal cuff pressure.