BACKGROUND: Studies showed cellular adhesion molecule and neurotrophic factor secreted from olfactory ensheathing cells (OECs) could protect the spinal neurons and promote the regeneration of spinal axon. OBJECTIVE: To compare the competence to repair spinal cord injury between olfactory mucosa OECs and olfactory bulb OECs. DESIGN, TIME AND SETTING: Randomized control animal experiment was performed in the central laboratory of Xidian Group Hospital between June 2007 and June 2008. MATERIALS: Twelve male SD rats were randomized selected and divided into experiment group (n=6, 23 months old) and control group (n=6, 3 months old). They were used for in vitro culture and purification of OECs; other 30 SD rats were randomized into three groups of 10 rats each: neonatal rat olfactory bulb OECs transplantation group, normal olfactory mucosa OECs transplantation group and blank control group.METHODS: Spinal cord injury models were produced in 30 rats, which were transplanted with the neonatal rat or SD rat OECs cultured in vitro. No transplant was given in the control group. MAIN OUTCOME MEASURES: At 4 and 8 weeks postoperation, the Basso, Beattie, Bresnahan (BBB) score for nerve function, the evoked potential of legs and the histopathological diversify of injured spinal cord. RESULTS: Seven rats died dudng the experiment process, and the death rate was similar between groups. At 4 and 8 weeks postoperation, there was no significant difference in the BBB scores between neonatal rat olfactory bulb OECs transplantation group and normal olfactory mucosa OECs transplantation group (P > 0.05), which were both significantly higher than blank control group (P < 0.001); the BBB scores in two transplantation groups were higher at 8 weeks than at 4 weeks (P < 0.01 ). At 4 weeks postoperaUon, no animal was shown to elicit motion evoked potential, but it was present in two transplantation groups at 8 weeks, with no significant difference between two groups (P > 0.05). The blank control group had still no motion evoked potential (P < 0.001 ). At 8 weeks postoperation, more cell infiltrations were found in the injured spinal cord of two transplantation groups, while few in the control group.CONCLUSION: Both OECs dissociated from olfactory bulb and olfactory mucosa have the same ability to repair the injured spinal cord, and their effect is similar.

Background Studies showed that inflammatory process participates in the pathogenesis anddevelopment of diabetic retinopathy targeting retinal vascular endothelial cells (RVECs).A growing body of evidence revealed that metformin reduces the risk of micro-and macro-vascular complications by protecting blood-brain barrier,however,whether it plays a protective effect on human retinal vascular by similar mechanism is still unelucidated.Objective This study was to investigate the effects of metformin on the proliferation,migration and secreting monocyte chemotactic protein-1 (MCP-1) and interleukin-8 (IL-8) of human retinal vascular endothelial cells (RVECs) under the stimulation of tumor necrosis factor-alpha (TNF-α).Methods RVECs were cultured and divided into normal control group,metformin (5 mmol/L) group,TNF-α 2.5 ng/ml group,and TNF-α+metformin (5,10,20 and 40 mmol/L,respectively) groups.Corresponding drugs were added into medium according to grouping for 24 hours.Cell numbers were calculated before and after treatment.The metabolic activity (absorbancy) of RVECs was measured with MTS assay.Cell migration of RVECs was assessed with transwell migration assay.The MCP-1 and IL-8 concentrations in the cell supernatant were detected by ELISA assay.Results The number of the cells was significantly different among the normal control group,metformin group,TNF-α group,and TNF-α+metformin (5,10,20 and 40 mmol/L,respectively) groups (F =189.31,P ＜ 0.01).The metabolic activities of RVECs were 0.32 + 0.02,0.32±0.03,0.97 ± 0.02,0.90 ± 0.05,0.76 ± 0.15,0.74 ± 0.05 and 0.41 ± 0.03;migrated cell numbers were (1 214±49),(1 200±45),(1 648±43),(1 309±48),(1 279±73),(961±60) and (942±106)/field;the concentrations of MCP-1 were (0.385 ±0.050),(0.362±0.060),(2.285 ±0.200),(1.131 ±0.180),(0.622 ± 0.120),(0.537±0.090) and (0.492±0.130) μg/ml,and those of IL-8 were (0.385±0.080),(0.390±0.120),(1.123±0.130),(0.899±0.180),(0.680±0.060),(0.417±0.090) and (0.335±0.100) μg/ml in the normal control group,metformin group,TNF-α group,and TNF-α + metformin (5,10,20 and 40 mmol/L,respectively) groups,showing significant differences among the groups (F =73.31,103.89,150.92,268.32,all at P＜ 0.01).The cell number,cell metabolic activity,migrated cell number,and MCP-1 and IL-8 levels in the cell supernatant were evidently increased in the TNF-α group compared with the normal control group,and those in the TNF-α+10 mmol/L metformin group,TNF-e +20 mmol/L metformin group and TNF-α+40 mmol/L metformin group were significantly decreased in comparison with the TNF-α group (all at P＜0.05).Conclusions Metformin can inhibit TNF-α-induced proliferation,migration and MCP-1 and IL-8 secretion of the cells,and therefore plays a protective role on RVECs in the inflammatory environment.

Objective To investigate the efficacy of transabdominal ultrasound interventional puncture combined with medicine in the treatment of ovarian chocolate cysts. Methods According to the different treatment methods will be January 2015 to December 2016 in Yuyao People's Hospital 70 cases of patients with ovarian chocolate cyst group: the control group with washed cysts + ethanol sclerotherapy by transabdominal ultrasound interventional puncture + urokinase solution, the observation group in the control group based on the addition of triptorelin;observed the body hormone before and after E2 two patients in the treatment group, follicle stimulating hormone, serum luteinizing hormone, FSH/LH, antral follicles level changes, complications, clinical treatment, and the relevant data for comparative analysis. Results Abdominal ultrasound interventional puncture and drug treatment (observation group) treatment of ovarian chocolate cyst is better than transabdominal ultrasound interventional treatment (control group), the stability of the body hormone levels of patients than the control group, the complication rate was lower than the control group, the total efficiency of clinical treatment was higher than the control group, the difference was statistically significant (P <0.05). Conclusion The patients with ovarian chocolate cyst transabdominal ultrasound interventional puncture and drug treatment effect significantly, can effectively improve the patients body hormone level, low complication rate, the total effective rate of the treatment is high, it is worthy of widely used treatment of patients with ovarian chocolate cyst.

Objective: To investigate the hydrolysis conversion rate of alcohol amine-diterpene alkaloids from aconitum alkaloids, hydrolyze aconitum alkaloids reference substance, calculate the amount of alcohol amine-diterpene alkaloids in the hydrolysis solution by the hydrolysis conversion rate, which is used as the amount of alcohol amine-diterpene alkaloids reference substance, and establish a content determination method for aconine, hypaconitine and aconine in Aconiti radix cocta.Methods: Through controlling the hydrolysis conditions of aconitine, hypaconitine and mesaconitine, aconine, hypaconitine and aconine were obtained.The determination was performed on an Agilent ZORBAX Extend-C18 RRHT(2.1 mm×50 mm,1.8 μm) column with the mobile phase consisting of methanol(A)-water(B containing 0.1% formic acid and 2.5 mmol·L-1 ammonium acetate) with gradient elution by HPLC-QTOF-MS.The flow rate was 0.21 ml·min-1.The column temperature was 30 ℃.MS instrument was equipped with an ESI+ ion source.Results: Under the hydrolysis conditions of this study, the conversion rate of aconine from aconitine was 99.64%;the conversion rate of hypaconitine from hypaconine was 99.94%;the conversion rate of mesaconitine from mesaconine was 99.57%.The HPLC-QTOF-MS methodological investigation showed the 3 kinds of alcohol amine-diterpene alkaloids were with good linearity (r>0.999 1).The RSD of the precision, repeatability and stability tests were less than 5%.The average recoveries were within the range of 99.43%-100.10%.Conclusion: The validated method is simple, specific, reliable and reproducible.In the absence of reference substance, it can be used for the quality control of the herbs of Aconitum L.species.

Objective:To develop a method of quantitative analysis of multi-components by single marker( QAMS) for nine kinds of alkaloids in Xiaohuoluo pills. Methods: An HPLC-QTOF-MS method with an Agilent ZORBAX Extend-C18 RRHT(2. 1 mm × 50 mm,1. 8 μm) column was applied. The flow rate was 0. 21 ml·min-1 . The column temperature was 30 ℃. The mobile phase was methanol (A)-water (B;containing 0. 1％ formic acid and 2. 5 mmol·L-1 ammonium acetate) with gradient elution. The aconitine was used as the internal standard, and the relative correction factor ( RCFs) of hypaconitine, mesaconitine, benzoylaconine, benzoyl-hypaconine, benzoylmesaconine, aconine, hypaconine and mesaconine was respectively established, and the reproducibility inspection on the RCF was performed. The contents of the other 8 kinds of aconitum alkaloids were calculated according to the RCF. At the same time, an external standard method ( ESM) was performed for the content determination of the nine alkaloids. The results of the two methods were compared. The feasibility and accuracy of the QAMS method were verified. Results:Within a certain range,the RCF of hypacontine,mesacontine, benzoylaconine, benzoylhypaconine, benzoyl mesaconine, aconine, hypaconine and mesaconine to aconitine was 1. 736,1. 979,1. 0471,0. 9242,1. 2901,1. 3078,1. 2859,and 1. 0948,respectively. The QAMS method was established for determi-ning alkaloids. There were no significant differences between the results of the QAMS method and those of the external standard method ( ESM) . Conclusion:With the validation of methodology, the method established in our study can be used for the content determina-tion of aconitine, hypaconitine, mesaconitine, benzoylaconine, benzoylhypaconine, benzoylmesaconine, aconine, hypaconine and mesaconine in xiaohuoluo pills.

Objective To investigate the effect of moldavica total flavone on pulmonary function and collagen type Ⅲ in lung tissue of ovalbumin-induced bronchial asthmatic rats. Methods A total of 60 SD rats were randomly divided into control group, model group, dexamethasone group, and moldavica total flavone low, medium and high dose group. Rat asthma model was established by ovalbumin challenge methods except the control group. The control and model group were intragastrically given distilled water, medicine groups were intragastrically given moldavica total flavone and dexamethasone for 30 d, once per day. After the last administration, the pulmonary function and airway responsiveness including breathing frequency (F), minute ventilation (MVb), peak inspiratory flow (PIFb), peak expiratory flow (PEFb) and enhanced pause (Penh) were measured by using non-invasive measurement system (Penh system, Buxco, USA). The collagen type Ⅲ in lung tissue were detected by ELISA. Results The moldavica total flavone at 180 and 360 mg/kg decreased F and Penh (P<0.05, P<0.01), increased MVb, PIFb and PEFb (P<0.05), and decreased collagen type Ⅲ in lung tissue (P<0.05, P<0.01). Conclusion Moldavica total flavone can relieve the airway hyperresponsiveness and remodeling, improve respiratory function of ovalbumin-induced bronchial asthmatic rats.

Objective To investigate the time-effect relationship and dose-effect relationship of the expression of adenovirus mediating Smad 7 gene regulated by irradiation via Egr-1 promoter in the lung of C57BL mice. Methods Recombinant adenovirus (AD.Egr-Smad 7) was made up through replication-defective adenovirus enclosed radio-inducible elements from the Egr-1 gene promoter and cDNA encoding Smad 7. Mice infected by intratracheal instillation with AD.Egr-Smad 7 were irradiated to their whole lungs after 24 hours. 324 mice were randomly divided into 6 groups, and the lungs were harvested at different time points following irradiation with single dose of 8?Gy to observe the time-effect relationship of Smad7 gene expression with different time intervals. 108 mice randomly divided into 3 groups were irradiated respectively by different single irradiation dose and the lungs were harvested 5 hours after radiation to evaluated the dose-effect relationship of Smad 7 gene expression with different radiation doses. The expression of exogenous Smad 7 was detected by Western blot analysis. Results The expression of adenovirus mediating Smad 7 gene regulated by Egr-1 promoter could be induced by radiation markedly as compared with the control groups(P

Objective Objective To study the effect of adenovirus mediating Smad 7 gene regulated by radiation via Egr-1 on the primary tumor and lung metastasis in C57BL mice implanted with Lewis lung cancer.Methods The radio-inducible elements from the Egr-1 gene promoter were inserted upstream to a cDNA encoding Smad7 and integrated into a replication-defective adenovirus to generate recombinant adenovirus(AD.Egr-Smad 7).270 mice implanted with Lewis lung cancer in the hind legs were used and the experiment was started when the transplanted tumor diameter reached 0.8 to 1.0cm.Then three investigations were undertaken,each demanding 90 mice implanted with Lewis lung cancer respectively.To each group,90 mice models were randomized into 3 groups: the normal control group;the NS control group;and the implanted AD.Egr-Smad 7 group.Every 6 mice in each group were irradiated by different single dose to study the following: 1.The maximal and minimal diameters of the tumor were recorded to observe the tumor growth tendency,the tumor growth delay and the mice survival time,2.The incidence of lung metastasis two weeks after the radiation was recorded.3.The incidence of lung metastasis when the tumor volume was four times as large as that at the beginning of radiation was recorded.Results The adenovirus mediating Smad 7 gene expression regulated by irradiation via Egr-1 in C57BL mice implanted with Lewis lung cancer was able to inhibit the progression of the primary tumor and prolong the survival of the mice significantly as compared with the control group(P0.05).Conclusions The gene expression of AD.Egr-Smad 7 regulated by radiation is not risky in promoting the local progression and distant metastasis of Lewis lung cancer in mice.On the other hand,the gene expression of AD.Egr-Smad 7 regulated by radiation could inhibit the progression of the primary tumor and prolong the survival time of the mice significantly.It is safe,to some extent,of using AD.Egr-Smad 7 to block the signal transduction of TGF-?locally as a novel strategy for gene therapy aiming at the prevention of radiation-induced lung

Objective To investigate the efficacy and safety of using stereotactic radiotherapy (SRT) technique for conducting re-therapy after thoracic tumor radiotherapy.Methods Thirty-eight patientswith SRT after receiving thoracic conventional radiotherapy (RT) in our hospital from July 2012 to November 2014 were selected.The treatment target area included the lung local primary lesion,recurrent lesions and lung metastasis tumor.Results Median dose of previous RT was 48 Gy (30-56 Gy).Median biologically equivalent effective dose (alpha/beta=10.0,BED10) of receiving SRT was 62 Gy (39-72 Gy).Median follow up time was 12.30 months;1-,2-year local progression-free survival (LPFS) was 76.32％ and 63.16 ％ respectively.Median recurrence-free (RFS) and overall survival (OS) were 13.20 months and 21.00 months respectively.Grade 2 and 3 pulmonary toxicity was 15.79 ％ and 7.89 ％ respectively.Other grade 2-4 toxicities adverse reactions included chest pain (15.79 ％),fatigue (18.42 ％) and skin lesion(2.63％).No grade 5 toxic injury occurred.Conclusion SRT can be safely and effectively used in the patients previously receiving thoracic RT.

Epigenetic mechanisms influence gene expression and function without modification of the base sequence of DNA and may generate a genetic phenotype.Epigenetic modifications include DNA methylation,histone modifications,and deployment of noncoding RNA.There is growing evidence that epigenetic mechanisms could play a crucial role in the development of diabetic retinopathy (DR).Molecular biological methods which could maintain mitochondrial homeostasis through the regulation of epigenetic mechanisms may prevent the development of DR.Epigenetic-related treatment modalities will become the new direction of targeted therapy for DR.