To observe the expression of N - cadherin and fibronectin in retinal pigment epithelium ( RPE) cells in vitro under high glucose conditions, furthermore, to explore the effects of high glucose on epithelial -mesenchymal transition (EMT) in RPE cells.
●METHODS: Human RPE (hRPE) cells were cultured in vitro. Containing a final concentration of 60mmol/ L glucose was used for high glucose treatment. The cells were divided into normal glucose group (5. 5mmol/ L, NG) and high glucose group (24, 48 and 72h) respectively. The expression of N - cadherin and fibronectin in hRPE cells were evaluated by immunofluorescence and real -time PCR.
●RESULTS:RPE cells became disorganized and swollen over time under high glucose conditions, especially in 72h subgroup. lmmunohistochemical analysis revealed that the expression of N - cadherin in RPE cells under high glucose conditions was decreased compared with that in the control group, while the expression of fibronectin was increased. Real - time PCR results showed that the expression of N - cadherin mRNA in high glucose group was decreased at 24h compared with that in the control group, and declined markedly at 72h ( F = 12. 252, P =0. 000). There were no significant differences between the control group and the high glucose group at 24h, while the differences between the control group and the high glucose group (48 and 72h) were significant respectively (P < 0. 05 ). Meanwhile, the expression of fibronectin mRNA in RPE cells was increased in high glucose group at 24h, and reached the peak at 72h (F = 50. 543, P = 0. 000). There were no significant differences between the control group and the high glucose group at 24h. Compared with the control group, the expression of fibronectin mRNA in hRPE cells was increased significantly in high glucose group (48 and 72h) respectively (P= 0. 000, P= 0. 000).
●CONCLUSlON: The expression of epithelium marker N-cadherin is down - regulated under high glucose conditions in hRPE cells in vitro. Meanwhile, the expression of mesenchymal maker fibronectin is induced and appeared to EMT changes. Results of this study will enrich our growing understanding in proliferative diabetic retinopathy and hopefully lead to novel insights for the pathogenesis and therapeutic treatments.

Objective To investigate the effects of output power,action time and radiofrequency(RF) needle on the cool-tip radiofrequency ablation(RFA) by experimental tools and to determine the value of ultrasonography in size evaluation of RFA zone.Methods The cool-tip RFA to fresh calf liver were monitored by ultrasound.The experiments by single electrode needle were performed with different combination of output power (80 W,120 W) and time (5 min,8 min,10 min).The cluster needle was used for assessment at 5 min with different output power(80 W,120 W).After the end of trial,the longitudinal specimens were cut open.The view and size of the ablation zone were recorded with naked eyes.The pathological changes displayed by optical microscope were recorded as well.Results The measurement of ablation zone with naked eyes showed with the ablation zone expanded with time in 80 W-power cases,but the pace of expansion slowed down,but in 120 W-power cases,expansion of the ablation zone was not obvious; the ablation zone in 120 W-power was bigger than that in 80 W-power at 5 min,their difference decreased with time,and the ablation zones were similar at 10 min.The cluster needle can produce ablation zone with lesser aspect ratio than that of single electrode needle,consequently similar to circle.Ultrasonic measurement of the ablation zone had real discrepancy.Most of longitudinal diameters were greater than the real ones,while in large ablation lesions,vertical diameters were often less than the real ones.Under optical microscope,no change could be found in shape and structure of the cells in ablation zone.Conclusion The output power and performing time have impact on ablation.The high-power output increased heat production as well as reduction of heat conduction.Compared with single electrode needle,the cluster needle produced ablation zone closer to real hepatic tumor,thus has more reliable effect to small hepatocellular carcinoma with diameter around 2 cm.The ultrasond has a great significance in RFA guidance,but it could not accurately define the border of ablation zone.

This study reported the analysis of plasma phospholipid metabolism of the rats and the pathological biomarkers between the type 2 diabetes model control group (MC) and the normal control group (NC). SD rats were randomly divided into 2 groups: NC and MC. To investigate state of plasma metabolite profiling in normal body, type 2 diabetes mellitus (T2DM) model group using UPLC-Q-TOF/MS which was used as analysis tool in this research. The compounds were identified by UPLC-Q-TOF/MS based on MS/MS fragment ions information, element composition in MassLynx 4.1 and the Lipid Maps database. The sign of two groups of samples in specific markers for screening was through a software package in R software (BioMark software). The results show that the pathological markers were mainly phosphatidylcholine (PC) and triglycerides (TG); the 2-acyl PC in the MC group was less more obviously than that in the NC group; high carbon number and high degree of unsaturation of the TG was reduced under the condition of type 2 diabetes. In the state of type 2 diabetes, metabolic changes occurred in rat plasma phospholipids obviously, which had a close relationship with the occurrence and development of T2DM.
Animals ; Biomarkers ; blood ; Chromatography, High Pressure Liquid ; Diabetes Mellitus, Experimental ; blood ; Diabetes Mellitus, Type 2 ; blood ; Metabolome ; Phospholipids ; blood ; Rats ; Rats, Sprague-Dawley ; Tandem Mass Spectrometry

BACKGROUND:Integrated Traditional Chinese and Western Medicine minimaly invasive treatment for halux valgus based on wrapped curtain method with “8”-shaped bandage and sub toe pad external fixation has been used for a long time in the clinic. This method abandons the internal implant fixationandexternal plaster fixation. After surgery, patients could take care of themselves. However, theactivity of the broken end may cause fracture nonunion, which once aroused scholars’ question. Recently, with the continuous improvement of foot biomechanics research, foot finite element model and applications become a reality.
OBJECTIVE:To evaluate thestability of osteotomy after the operation of wrapped curtain method with“8”-shaped bandage and sub toe pad external fixation on the basis of static finite element method.
METHODS:A young female volunteer with halux valgus was selected, whose body weight was 58 kg, and right foot halux abductor valgus angle was 24°; intermetatarsal angle was 13°; proximal articulator set angle was 7°; distal articulator set angle was 7°. CT was used to scan the right foot. ABAQUS software was applied to establish a finite element model of right foot halux valgus bone, and model of the first metatarsal neck minimaly invasive osteotomy was simulated based on wrapped curtain method with external fixation. Von Mises stress and displacement at the osteotomy endwere calculated.
RESULTS AND CONCLUSION:(1) The maximum stress was 0.067 MPa without external fixation, and the maximum stress was 1.258 MPa with the external fixation. Stress was mainly distributed in the outer edge of the osteotomy. (2) The maximum absolute displacement was 0.363 mm without external fixation, and the maximum absolute displacement was 0.716 mm with external fixation. The two largest displacements were both in the Z-axis direction. Statistical analysis confirmed that the four nodes absolute displacement and stress were significantly different (P< 0.01). (3) The maximum relative displacement was 0.101 mm. The maximum relative displacement was 0.046 mm with external fixation. The maximum relative displacement without external fixation was-0.102 mm and occurred in the Z-axis. The maximum relative displacement with external fixation was 0.110 mm and occurred in the Y-axis. (4) One-way analysis of variance confirmed that the four nodes relative displacements were not statisticaly significant in X-axisand Y-axis (P> 0.05). The four nodes relative displacements were statisticaly significant in Z-axis (P< 0.05). (5) These findings suggest that the external fixation based on wrapped curtain method after halux valgus surgery could effectively reduce osteotomy displacement. The moderate stress and elastic fixation are conducive to fracture healing.

OBJECTIVE: To analyze risk factors associated with post-traumatic retinal detachment and to identify the cause-effect relationship between retinal detachment and blunt ocular trauma in forensic medical assessment. METHODS: 112 cases of forensic medical examination on post -traumatic retinal detachment were retrospectively analyzed. RESULTS A variety of retinal abnormalities were found in these cases, including tearing of the ora serrata retinae (4.28%), macular perforation (12.50%), small (<90 degrees) nonmacular retinal perforation (56.25%), larger (>90 degrees) retinal perforation (5.00%), and tractive retinal detachment without perforation (11.6%). Proliferative vitreoretinopathy (PVR) was found in 45 eyes examined. In most cases, retinal detachment occurred between 1 week to 2 months after blunt ocular trauma (61.60%), with 83.93% accompanied with severe myopia (>-3.00D) and 52.67% accompanied with vitreous floaters. Of all cases, 41.07% were directly and 52.68% were indirectly resulted from blunt ocular trauma, and the rest (6.25%) showed no association with blunt ocular trauma. CONCLUSION Many risk factors may result in retinal detachment including blunt ocular trauma and other causes. Accurate assessment of the relationship between blunt ocular trauma and retinal detachment is an important part of forensic examination.
Adolescent ; Adult ; Contusions/complications* ; Eye Injuries/complications* ; Female ; Forensic Medicine/methods* ; Humans ; Male ; Middle Aged ; Retinal Detachment/pathology* ; Retrospective Studies ; Time Factors ; Wounds, Nonpenetrating/complications* ; Young Adult

Abstract: Nitrobenzene compounds (NBCs) are widely used in the world. It has 40 isomers such as nitrobenzene, dinitrobenzene and nitrotoluene, that are highly toxic and difficult to degrade and can cause harm to human health in different degrees. At pres⁃ ent, there is no unified standard method and occupational exposure limit for the detection of NBCs in the air. In terms of sampling medium, solid adsorption tube is mostly used for trapping vapor state NBCs, and filter membrane and solid adsorption tube are mostly used in series for sampling coexist NBCs in vapor state and aerosol state. In the detection methods, gas chromatography and liquid chromatography are common, and ultraviolet spectrophotometry, Raman spectroscopy, ion migration spectrometry and some other rapid response methods and technologies are also used in the detection of NBCs. In the detection of NBCs by gas chro⁃ matography, capillary column separation is commonly used, and the main detectors are flame ionization detector, electron capture detector and mass spectrometry detector. It is of practical significance to establish a method with high sensitivity, strong practica⁃ bility, convenient operation, and can simultaneously collect and detect a variety of NBCs in different states.

Objective - - To prepare the GDH 5 air sampling tube for simultaneous collection of eight kinds of chloro nitrobenzene （ ） ， compounds CNBs in the air of workplace and establish a matching determination method using gas chromatography. Methods - - ， Eight kinds of CNBs in vapor and aerosol state were collected by self developed GDH 5 air sampling tube desorbed ， ， ， by toluene separated by polysiloxane gas chromatography column detected by microcell electron capture detector and Results - （ - quantified by external standard method. It was determined that the air sampling tube was assembled by XAD 2 ion ） - ， exchange resin and glass fiber filter membrane. The linear range of CNBs was 0.80 240.00 mg/L and the linear correlation - - coefficients were greater than 0.999 9. The detection limit was 7.87 13.03 μg/L. The minimum detectable concentration was 0.60 3， - 3（ ） 1.33 μg/m and the minimum quantitative concentration was 2.00 4.22 μg/m sample 45.00 L . The average desorption - - （RSD） - ， - RSD efficiency was 101.2% 110.0%. The within run relative standard deviation was 0.8% 4.1% and the between run - Conclusion - was 0.3% 5.8%. The samples could be stored for more than 30 days at room temperature. GDH 5 air sampling tube and its associated determination method can be used for the collection and determination of eight kinds of CNBs in workplace air.

Objective To explore the feasibility and advantages of establishment of retrograde amnesia (RA) model in mice with such methods as electric shock,anoxia,and anesthesia.Methods Kunming mice were divided into control group and 5 treatment groups,including treatments with electric shock,anoxia,propofol,electric shock+anoxia,electric shock+propofol.At first,every group received the same dark avoidance training to establish the behavior of dark avoidance,then the 5 treatment groups were treated with the methods of 120-180 V electric shock,anoxia within a closed container,intraperitoneal injection of 0.3 mL of propofol,electric shock+anoxia,electric shock+propofol,respectively.Next day,step-in latency (Lat) of mousse were measured with the dark chamber in all groups and changes of dark avoidance behavior were analyzed.Results The Lat in the control group 24 h after dark avoidance training was (111.7+17.2) s.In the treatment groups of electric shock,anoxia,electric shock+anoxia,electric shock+propofol,significantly shortened Lat,which limited to 30 s in some mice 24 hatter corresponding treatment,was observed as compared with that in the control group (P<0.05).Except for the propofol treatment group did not changed obviously,the incidence rate of shortened Lat was 43.8%,45.4%,66.7% and 60% in the electric shock treatment group,anoxia treatment group,electric shock+anoxia treatment group,and electric shock+propofol treatment group,respectively.On the 5th and 8th d,some mice recovered from the shortened Lat.Conclusion RA model can be established successfully in some mice treated with electric shock,anoxia,electric shock+anoxia,electric shock+propofol and the highest modeling rate was found in the electric shock+anoxia treatment group.RA can recover in the later stage in some modeling mice and use of pmplfol alone call not induce RA.

The Notch signaling pathway has been implicated in the regulation of cell-fate decisions such as differentiation of embryo stem cells and neural stem cells into neurons. We cultured human mesenchymal stem cells (hMSCs) in vitro and induced hMSCs to differentiate into neural cells by beta-mercaptoethanol (beta-ME), DMSO and 3-tert-butyl-4-hydroxyanisole (BHA). Immunocytochemistry was utilized to detect neuron-specific enolase (NSE) and Nissl body, and flow cytometry was used to determine cell growth phases. The expressions of signal molecules involved in the Notch pathway such as Notch1, Jagged 1 (JAG1), presenilin 1 (PS1) and hairy and enhancer of split 1(HES1) were observed by RT-PCR and immunofluorescent techniques. The results were as follows: (1) Before induction, the percentage of hMSCs at G(0)/G(1) was 58.5%, and the percentage at S+G(2)/M was 41.5%. After induction, the percentage of hMSCs at G(0)/G(1) increased to 73.1%, 76.2% and 78.1%, respectively on days 2, 4 and 6, and the percentage at S+G(2)/M decreased to 26.8%, 24.8% and 21.9%, respectively; The percentage of NSE-positive cells reached (77+/-0.35) %; Nisslos staining was positive in cytoplasm. (2) Notch1 and JAG1 were both expressed in hMSCs before and after induction, but the mRNA expressions of both Notch1 and JAG1, detected by RT-PCR, decreased obviously after induction(P<0.05). Notch1 mRNA/beta-actin was 1.157, 0.815, 0.756 and 0.570, and JAG1 mRNA/beta-actin was 0.437, 0.350, 0.314 and 0.362, respectively, on days 0, 2, 4 and 6 after induction. The Notch pathway activation participant PS1 mRNA and Notch pathway target gene HES1 mRNA also decreased apparently after induction (P<0.05), and their mRNA/beta-actin was 0.990, 0.449, 0.441, 0.454 and 0.370, 0.256, 0.266, 0.240 on days 0, 2, 4 and 6, respectively. These observations indicate that the expressions of Notch signal molecules were suppressed when hMSCs were induced to differentiate into neural cells. Based on these findings, we propose that low level of Notch signaling activation may contribute to neural cell differentiation.
Basic Helix-Loop-Helix Transcription Factors ; genetics ; Calcium-Binding Proteins ; genetics ; Cell Cycle ; Cell Differentiation ; Flow Cytometry ; Homeodomain Proteins ; genetics ; Humans ; Intercellular Signaling Peptides and Proteins ; genetics ; Jagged-1 Protein ; Membrane Proteins ; genetics ; Mesenchymal Stromal Cells ; cytology ; Neurons ; cytology ; Receptor, Notch1 ; genetics ; Receptors, Notch ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Serrate-Jagged Proteins ; Signal Transduction ; Transcription Factor HES-1

OBJECTIVETo investigate the effect of platelet-rich fibrin (PRF) on human gingival fibroblasts (HGF) biological behavior such as proliferation, migration and collagen-I expression.

METHODSHuman healthy gingival tissues were cultured according to the explant technique to obtain primary cultures. PRF was prepared by means of Choukroun's. HGF were co-cultured with PRF membrane originating from the same donor as the explants, divided into three groups, PRF1 group, PRF2 group and blank control group. Methyl thiazolyl tetrazolium (MTT) assay was used for cytotoxicity and cell proliferation study, and enzyme-linked immunosorbent assay (ELISA) for collagen-I (COL-I) secretion study at the 1st, 3rd, 5th day respectively. Eluates from PRF membrane was prepared, and divided into three groups, PRF1 group, PRF2 group and blank control group. Transwell chamber was utilized to determine the effect of PRF membrane eluate on cell migration.

RESULTSThe A values of HGF in culture of the PRF1 (0.615 ± 0.036, 0.686 ± 0.006, 0.693 ± 0.004) and PRF2 groups (0.653 ± 0.023, 0.766 ± 0.034, 0.775 ± 0.053) were significantly higher than those of the control cultures (0.514 ± 0.020, 0.544 ± 0.006, 0.545 ± 0.009) (P < 0.01), but the difference between PRF1 and PRF2 group was not significant (P > 0.05). In each group at different time points, the HGF proliferation effect was significantly enhanced with time (P < 0.01). Cell migration test showed that the migration numbers of HGF in PRF1 and PRF2 groups (85.67 ± 2.94, 85.83 ± 1.47) were significantly higher than those of the control group (54.17 ± 2.48) (P < 0.01), but the difference between the two experimental groups was not significant (P > 0.05). COL-I secretion test exhibited that the A values of COL-I in PRF1 (0.184 ± 0.004, 0.200 ± 0.004, 0.204 ± 0.009) and PRF2 group (0.213 ± 0.008, 0.226 ± 0.005, 0.229 ± 0.006) were significantly higher than the A values of the control group (0.174 ± 0.002, 0.184 ± 0.002, 0.186 ± 0.003) (P < 0.01), but the difference between the two experimental groups was not significant (P > 0.05). In each group, the secretion level of COL-I increased significantly with time (P < 0.01).

CONCLUSIONSPRF could exert a positive effect on HGF biological behaviour and had clinical application potential in the treatment of gingival recession and in the periodontal tissue engineering when combined with seed cell HGF.

Adult ; Blood Platelets ; Cell Movement ; Cell Proliferation ; Collagen Type I ; metabolism ; Fibrin ; pharmacology ; Fibroblasts ; cytology ; secretion ; Gingiva ; cytology ; Humans ; Male ; Primary Cell Culture