BACKGROUND: Human pulp tissue has been known to be less, and exhibit poor tolerance to enzymatic digestion and less adherent cells after step-by-step digestion of trypsin and collagenase, thereby often leading to a failure of passage. Only several kinds of dental pulp cells with poor activity can be obtained by the tissue explant-collagenase digestion. OBJECTIVE: To investigate human dental pulp cells cultured in vitro by tissue explant method. DESIGN, TIME AND SETTING: A cytological observation was performed at Heping Campus and School of Stomatology, Jilin University from 2005 to 2007. MATERIALS: Healthy young human teeth extracted for orthodontic correction or impaction. METHODS: Pulp tissue from the third molar teeth was collected, cut into small blocks with a size of 1.0 mm×1.0 mm×0.5 mm under the infiltration of small amount of Dulbecco's modified eagle's medium, and then transferred into a 6-well plate containing culture medium for incubation in a 5% CO2 and saturated humidity atmosphere at 37 ℃. During the process of incubation, pulp tissue was adjusted at a density of 3-6 blocks/well, with an equal spacing of 0.5 cm and the 6-well plate was kept inverted. Three hours later, the 6-well plate was turned over to make tissue blocks adhering to the plate wall. Culture was continued after addition of 2 mL of culture medium. Culture medium was renewed every 4-6 days. After 6-15 days, cells emigrated from the edge of tissue blocks and call outgrowth appeared around each tissue block. When cells closed to confluency, a digestion procedure of 2.0-3.0 minutes (0.25% trypsin and 0.02% ethylenadiamine tetraacetic acid) was followed by passage culture at a proportion of 1: (2-3) in 25 mL of culture flasks. Purified fibroblast-like cells were gradually obtained from primarily cultured cells by repeated digestion and passage. MAIN OUTCOME MEASURES: Cellular morphology was identified by immunohistochemistry; secreted dental pulp cells were determined using alkaline phosphatase activity; the growth curves of human pulp tissue cells were depicted by MTT assay. RESULTS: Under an inverted phase contrast microscope, the obtained dental pulp cells were primarily typical fibroblasts with a long-shuttled appearance, well-rounded call body, uniform cytoplasm, round or oval nucleus, and clear nucleolus. Immunohistochemistry results showed call surface vimentin-positive, pan cytokeratin-negative, and alkaline phosphatase-posltive These cells were decreased after culturing 1 day, were slightly increased after 2 days, entered the logarithmic growth period and were markedly increased after 4 days, entered a platform period after 8 days, and began to decrease again after 9 days. The whole growth curve of cells appeared in "S" shape.CONCLUSION: The dental pulp cells isolated from human pulp tissue by tissue explant method can effectively proliferate end retain a poody differentiated state in vitro.

Objective To investigate the effect of different concenrtrations of basic fibroblast growth factor (bFGF) on proliferation of human dental pulp cell in vitro,and to find out the most effective concentration of bFGF. Methods Dental pulp cells were isolated from dental pulp tissue explants.The pulp cells were divided into 5 groups randomly,bFGF was added into each group until the ultimately concentrations were 0.1,1.0,10.0 and 100.0 ?g?L-1respectively while the group without bFGF as control group. The effects of bFGF on dental pulp cells were assayed by absorbency A and relative growth rate(RGR) with MTT colorimetric method. Results bFGF at concentrations of 1.0-100.0 ?g?L-1 promoted the cell proliferation (P0.05). Conclusion bFGF has the capability of promoting the proliferation of human dental pulp cells,and the smallest effective concentration is 1.0 ?g?L-1,the most strong cell proliferation takes place at bFGF concentration of 10.0 ?g?L-1.

Objective To explore the effect of flexible laser endotracheal intubation optical device (patent number 201520785723.7) on oral and maxillofacial intubation operations. Methods Forty-two patients with the oral and maxillofacial intubation optical device for general anesthesia were included in this study. The patients were from 16 to 73 years old and ASAⅠ-Ⅱ. Surface anesthesia was prepared for airway and nasal cavity. There were 27 patients whose mouth opening were smaller than 3 cm. After intravenous injection of midazolam 0.05 mg/kg and sufentanil 0.2μg/kg, reinforced endotracheal tube was inserted into throat 13-15 cm, then the probe shape to 120° and optical device were placed into the tube respectively, guided the tube according to the light spot in front of neck. There were 15 patients whose mouth opening were not smaller than 3 cm. The quick guidance optical device and larygoscope were adopted to help the intubation. After intravenous injection of midazolam 0.05 mg/kg, sufentanil 0.5 μg/kg, propofol 2 mg/kg, cisatracurium 0.2 mg/kg, the reinforced endotracheal tube was inserted into the oral cavity nasally, and completed intubation with laryngoscope. The success rate of intubation, time of intubation, the real-time blood pressure before and after intubation, heart rate and related complications were recorded before intubation and after intubation. Results In the 42 patients, 41 patients were successful for guidance intubation, and the success rate of intubation was about 97.6%. The average intubation time was(124.5±38.2) seconds. The shortest intubation time was 12 seconds and the longest was 276 seconds. During intubation, the increases of blood pressure and heart rate were in the normal range (<30%). Levels of SpO2 were maintained at 0.92-1.00. All the patients were without agitation. There were no serious complications such as hoarseness and pharyngalgia after operation.Conclusion The flexible laser endotracheal intubation optical device can be used quickly for oral and maxillofacial intubation, and the 45° bevel design for optical device can form two kinds of zones with bright light upward and downward. It is a new tool for nasal intubation that is more convenient for location of light guidance, and has high success rate and small stimulation.

Objective To evaluate the value and feasibility of Microsim medical simulation training system in medical students' clinical thinking training. Method 96 students of 5-year program of medicine of Grade 2009 and Grade 2010 were the research object. These students were randomly divided into two groups (group A:After 3 weeks' clinical practice in respiratory medicine, taking 1 week Microsim training. group B: Taking 4 weeks clinical practice in respiratory medicine. Each group has 48 students.). The examination and teaching satisfaction of the two groups were observed after the end of the internship. SPSS 17.0 statistical software was used to analyze the collected data (measurement data matching t test, counting data by chi-square test). Results The Microsim system score: group A was (89.37±7.18), group B was (61.95±15.34). The difference between groups was statistically signifi-cant. The following scores suggested the assessment of students' ability of clinical thinking: ability to analyze problems [group A (89.95±4.02) vs. group B (75.51±6.34)], the ability to deal with the prob-lem [group A (78.81±8.09) vs. group B (59.67±9.33)], treatment scheme [group A (86.74±6.59) vs. group B (70.39±7.05)] and the treatment effect [group A (88.61±8.16) vs. group B (63.54±11.48)]. In these aspects, the two groups had statistically significant difference, but communication [group A (82.47 ±5.23) vs. group B (84.09 ±3.72)] had no statistically significant difference between the two groups. 89.6% (43) of the participants believed that the Microsim medical simulation training system could significantly improve the clinical thinking ability, but only 58.3% (28) of the students believed that the basic theory of knowledge could be consolidated. Conclusion Microsim medical simulation training system can improve the students' ability of clinical thinking and clinical comprehensive treat-ment ability. It can be used as an effective complement to clinical practice teaching.

Objective:To explore the impact of compound nutrients supplementation on nutritional and immune status in middle-aged and elderly people.Method:The volunteers aged 50-65 years old were randomized into two groups.The supplement was given to the test group,the control group took placebo.Results:In comparison with the control group,the test group had a higher level of serum vitamin A and zinc,blood selenium.The humoral immune indices as IgA、IgG、IgM、C3 of the test group were higher than those of the control group,but no significant difference of cellular immunity indices between two groups.Conclusion:Supplementation of compound nutrients could improve the nutritional status of vitamin A,zinc and selenium,and may promote humoral immunity.

Objective To evaluated the risk factors of urethral recurrence ( UR) following radical cystectomy ( RC) in patients with bladder urothelial carcinoma.Methods The clinical data of 350 male patients who underwent RC between January 2005 and January 2013 were retrospectively analyzed.The mean age was 63 years (rang 46-76) years.176 cases had the history of non-muscle-invasive bladder cancer.15 cases were were found the tumor invasion into the prostatic urethral.The way of urinary diversion after RC included 172 cases were orthotopic neobladder, 90 cases were cutaneous diversion and 88 cases were ileal couduitin.331 cases underwent preoperation intravesical instillation.36 cases underwent systemic chemotherapy after operation.148 cases were found the multiple tumor lesions, which was more than 2 sites. The pathological stage was more than T2 satge in 189 cases.And 177 cases were diagnosed as high-grade urothelial carcinoma.Multivariate Cox regression analyses were used to evaluate the risk factors associated with the UR.Results There were 350 cases in this study, UR was observed in 28 cases ( 8%).On multivariate Cox regression analyses, previous history of NMIBC (HR=15.205,95%CI 3.718-62.180,P<0.001), prostate urethral involvement(HR=5.233,95%CI 1.106-24.754,P=0.037) and Non-orthotopic neobladder(HR=6.656,95%CI 1.840-24.077,P=0.004)which the operation of cutaneous diversion and ileal couduit , were independent risk factors of UR following RC.Intravesical instillation before operation ( HR =0.470, 95%CI 0.010-0.217, P <0.001 ) was the protective factor of the UR.Conclusions Previous history of NMIBC, prostatic urethral involvement and Non-orthotopic neobladder were independent risk factors of UR.Intravesical instillation before operation was protective factor of UR.Urethrectomy for patients with high risk factors and intravesical instillation before operation were important.

AIM:To dynamically observe and compare the relative changes of the indexes from the process of acute inflammation to chronic remodeling in asthmatic mice induced by ovalbumin ( OVA) .METHODS:Female BALB/c mice (n=60) were randomly divided into normal control group and asthma group .The mice in asthma group were sensi-tized and challenged by OVA , while the mice in normal group received equal volume of normal saline ( NS) .The challenge was performed for 3 consecutive days from the 21th day to observe the response of acute inflammation , and then the mice in different groups were challenged once per week for 5 weeks.Detailed comparisons of the dynamic changes of cell infiltra-tion, cytokine expression and airway remodeling were conducted .RESULTS:Compared with NS group , the mice in OVA group showed a predominantly eosinophilic infiltration into the airway lumen , increased production of Th 2-type cytokines , secretion of epithelial mucus and deposition of subepithelial collagen .In OVA challenge groups , the levels of inflammatory cells and inflammatory factors were remarkably higher in 24 d group, whereas the most obvious changes of goblet cell hyper-plasia and airway remodeling were observed in 52 d group.CONCLUSION:Acute asthma model is sufficiently induced by 3 consecutive days of OVA challenge protocol , which is accompanied with high levels of inflammatory cells and inflammato-ry factors.The OVA challenge protocol once per week for 5 weeks could induce a chronic asthma model with obvious airway remodeling .

Objective To investigate the expressions and the relationship of FEZ1 and p16 in cervical intraepithelial neoplasia (CIN).Methods The expressions of FEZ1 and p16 in 93 cases of CIN and 10 cases of normal cervical specimens were detected by immunohistochemistry.Results The positive expression rates of FEZ 1 were 90.0％ (9/10) of normal cervical specimens,67.9％ (19/28) of CIN I,36.0％ (9/25) of CIN Ⅱ,22.5％(9/40) of CIN Ⅲ.The positive expression rates of p16 were 10.0％(1/10) of normal cervical specimens,32.1％(9/28) of CIN I,76.0％(19/25) of CIN Ⅱ,92.5％(37/40) of CIN Ⅲ.There were significant differences in the positive expression rates of FEZ1 and p16 between CIN Ⅰ and CIN Ⅲ (P ＜ 0.01),there was no significant difference in the positive expression rates of FEZ1 and p16 between CIN Ⅱ and CIN Ⅲ.The expression of FEZ1 and p16 was negatively correlation in CIN (r =-0.712,P＜ 0.01).Conclusion The abnormal high expression of p16 and abnormal low expression of FEZ1 in CIN may be involved in the occurrence and development of CIN,detecting the expressions of the two indexes may be helpful for clinical diagnosis.

BACKGROUND:Previous studies have demonstrated that transplantation of control ed release glial cellline-derived neurotrophic factor and bone marrow mesenchymal stem cells-derived neuron-like cells can effectively promote the motor function and sensory function recovery of rhesus monkeys with spinal cord injury.
OBJECTIVE:To validate whether co-transplantation of control ed release glial cellline-derived neurotrophic factor and bone marrow mesenchymal stem cells-derived neuron-like cells exhibits better protective effects on spinal cord glial scar of rhesus monkeys with spinal cord injury than celltransplantation alone.
METHODS:Twelve rhesus monkeys were col ected to prepare animal models of acute severe spinal cord injury using modified Al en’s method, and then randomly divided into three groups:experimental group, co-transplantation of control ed release glial cellline-derived neurotrophic factor and bone marrow mesenchymal stem cells-derived neuron-like cells;control group, simple transplantation of bone marrow mesenchymal stem cells-derived neuron-like cells;blank control group, PBS. After 5 months, paraffin specimens of the spinal cord were made for detection of morphological and compositional characteristics of glial scar, regeneration of nerve fibers in the scar, glial scar area, and average absorbance of glial fibril ary acidic protein.
RESULTS AND CONCLUSION:Glial scar in the injured spinal cord was composed of astrocytes and histocytes. Less spinal cord glial scar area and lower absorbance value could be observed in the experimental and control groups as compared with the blank control group (P<0.05). In addition, in the blank control group, neurofilament negative fibers could be observed in the glial scar, and astrocytes proliferated obviously. The experimental and control groups showed less fibers passed through the scar area. The glial scar area and average absorbance in the experimental group was lower than that in the control group (P<0.05). These findings suggest that compared with simple transplantation of bone marrow mesenchymal stem cells-derived neuron-like cells, co-transplantation of control ed release glial cellline-derived neurotrophic factor and bone marrow mesenchymal stem cells-derived neuron-like cells shows better protective effects spinal tissue structure after spinal cord injury, which may be one of mechanisms by which the number of glial scars is reduced to a greater extent.

Objective To explore the effect and mechanism of Kuntai Capsule (KC) on premature ovarian failure in rats.Methods Totally 40 SD female rats were randomly divided into four groups:control group,model group,Kuntai Capsule (KC) group and conjugated estrogens tablets (CET) group.The premature ovarian failure model in rats was made by ig administration of 75 mg/kg tripterygium.After the model was established,rats were continually treated with 0.6 and 0.625 g/kg of KC and CET respectively by ig administration for 36 d.HE staining was used to observe the morphology of ovary and count the number of follicle.The enzyme-linked immunosorbent assay (Elisa) method was used for the detection of follicle stimulating hormone (FSH),luteinizing hormone (LH),serum estradiol (E2),and anti-Mueller tube hormone (AMH) level.The real-time fluorescence quantitative PCR (qRT-PCR) method was used to detect the mRNA expression of vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF).Results Compared with control group,the number of primordial follicles,preantral follicles and antral follicles were significantly reduced,the number of atretic follicles was increased (P ＜ 0.05),E2 and AMH significantly decreased (P ＜ 0.05),FSH and LH levels increased significantly (P ＜ 0.05),and the levels of VEGF and bFGF mRNA in model group were significantly decreased (P ＜ 0.05).Compared with model group,the number of primordial follicles,preantral follicles,and antral follicles were significantly increased,the number of atretic follicles was significantly reduced (P ＜ 0.05),E2 and AMH significantly increased (P ＜ 0.05),FSH and LH levels significantly decreased (P ＜ 0.05),and the levels of VEGF and bFGF mRNA in KC group were significantly increased (P ＜ 0.05).Conclusion By regulating the level of hormone and up-regulating the expression of VEGF and bFGF,KC can repair the damaged ovarian tissue and promote the growth and development of the follicle,so as to inhibit the premature exhaustion of mRNA.