Objective To investigate the expression of microRNA-34a (miR-34a) and silent information regulator 1 (SIRT1) in human lens epithelial cells under H2O2-induced oxidative stress.Methods Different concentrations of H2O2 (0 μmol · L-1,100 μ mol· L-1,200 μmol · L-1,300 μmol · L-1,and 400 μmol · L-1) were used to stimulate SRA01/04 cells for 24 hours.Cell viability was measured using cell counting kit-8 (CCK-8) assay.Cell apoptosis was detected by flow cytometry.Expression levels of miR-34a/SIRT1 were measured by RT-PCR.Results CCK-8 assay showed that a certain concentration range of H2O2 had a proliferation inhibition on SRA01/04 cells.There was a dose response relationship between 100 μmol · L-1 and 400 μmol · L-1.Compared with 0 μmol · L-1 H2O2 group,the difference was statistically significant (all P ＜ 0.01).According to flow cytometry results,apoptotic rate of SRA01/04 cells in control group and H2O2(100-300 μmol · L-1) groups were (6.1 ± 1.2)％,(26.3 ± 1.8)％,(32.5 ± 2.2) ％,and (64.7 ± 5.3) ％.Compared with 0 μmol · L-1 H2 O2 group,the differences were statistically significant (all P ＜ 0.01).RT-PCR test results showed that the expression of miR-34a increased significantly in a dose-dependent manner after the SRA01/04 cells treated with different concentrations of H2O2,while SIRT1 expression level was decreased,there were significant differences compared with control group (all P ＜ 0.001).Conclusion There is a significantly increase of miR-34a and decrease of SIRT1 in human lens epithelial cells under the oxidative stress of a certain concentration of H2O2.Down-regulated expression of miR-34a can increase the survival rate of human lens epithelial cells under H2O2-induced oxidative stress.

Background Corneal blindness is one of the major blinding eye diseases in China.With the development and progress of tissue engineering technology,tissue-engineered corneas offers a new approach to the treatment of corneal diseases.To select and cultivate ideal seed cells is a foundation of construction of tissueengineered corneas.Objective This study was to evaluate the efficiency of stripe off the Descemet membrane with endothelium plus enzymic digestion in the isolation of corneal endothelial cells and analyze the bionomics of rabbit corneal endothelial cells (CECs) in vitro.Methods Descemet membrane was stripped from fresh cornea of New Zealand rabbit under the dissection microscope.Descemet membrane with endothelium was incubated in trypsin and EDTA solution at 37 ℃ and then purified for CECs subculture in vitro.The morphology of the cultured cells was observed under the inverted microscope and marked by CM-Dil dye solution.Then the shape of the cells was observed by hematoxylin and eosin staining and the cells were identified for the expression of neuron specific enolase (NSE) using immunochemistry.The viability of the cells were evaluated by trypan blue staining.The surface structure of the cells were examined under the scanning electron microscope.Intercellular zonula occludens-1 (ZO-1) was identified by immunofluorecsence staining.Results A large number of purified CECs were obtained from Descemet membrane with endothelium through enzymic digestion.Cultured cells grew well and formed monolayer 5-7 days later with the cobblestone stone-like arrangement.The survival rate of the cells was 95％.CECs presented with the red annular fluorescence for CM-Dil with the labeling rate ＞90％.NSE was positively expressed in the cytoplasm.Polygon CECs were seen by hematoxylin and eosin staining and showed the brown staining.Abundant microvilli on the cellular surface and interconnected foot process were seen under the scanning electron microscope.ZO-1 showed the green fluorescence.Conclusions The method of striping off the corneal Descemet membrane with endothelium plus enzymic digestion can obtain abundant CECs.Cultured cells have good biological properties.This study may offer a feasible application in the engineering of corneal transplant membrane.

Gastric cancer incidence is one of the most common malignancies in our country and is the second most common in the worldwide,clinic doctors always emphasize early diagnosis and treatment of gastric cancer patients,in order to reduce the mortality,however,most patients' condition often have been in the late fall and these patients were badly in efficacy.Looking for a new diagnosis way is a medical prddem,with molecular biology advance and gene chips was improved,it is possible for the early screening of gastric cancer.This assay aims to briefly analyse the role of gene chips in the research progress of early gastric cancer.

Objective To evaluate whether there is a correlation among plasma levels of procalcitonin (PCT),N-terminal pro-brain natriuretic peptide (NT-pro-BNP)and cardiac troponin T (cTnT)in patients with sepsis,as well as significance to prognosis of patients.Methods 48 patients with sepsis who were admitted to the intensive care unit of a hospital between September 2014 and March 2015 were chosen for study,patients were divided into severe and mild sepsis groups according to the disease condition,plasma levels of PCT,NT-pro-BNP,and cTnT were de-tected,mortality of patients were analyzed statistically,relation between plasma levels of PCT,NT-pro-BNP,cT-nT and patients’death were compared.Results The plasma levels of PCT,NT-pro-BNP and cTnT in severe sepsis group were significantly higher than those in mild sepsis group (all P <0.05);mortality of mild sepsis group was significantly lower than that of severe sepsis group (10.53% vs 41.38%,P <0.05);Levels of PCT,NT-pro-BNP and cTnT levels in died patients were all higher than surviving patients (all P <0.05);levels of PCT and NT-pro-BNP,NT-pro-BNP and cTnT were positively correlated respectively (rs = 0.337,P <0.05;rs =0.456,P =0.001, respectively ),while PCT was not significantly correlated with cTnT.Plasma levels of PCT,NT-pro-BNP,and cTnT were all correlated with the prognosis of patients (P <0.05),and is helpful for judging the prognosis of pa-tients,combination of three indexes had better prognostic value for the prognosis.Conclusion Combination detec-tion of plasmid levels of PCT,NP-pro-BNP,and cTnT can assess the severity of infection in patients with sepsis, and preliminarily judge the prognosis of patients with sepsis.

Objective To evaluate the clinical effect of the cannulated screw fixation in treatment of the dens fracture under locational guidance. Methods The study involved 27 patients treated with the cannulated screw fixation under locational guidance from January 2005 to January 2009.There were 19 patients with type II fracture and eight with type light m fracture.The lateral and open mouth position X-ray examination of the cervical ventebrae was done to observe the fracture healing. Results The operation lasted for a range of 40 minutes to 1.3 hour (average 1.0 hour),which showed no any complications.The patients were followed up for average 6.5 months(3-12 months),which showed sound fracture healing in 26 patients but nonhealing in one. Conclusion The cannulated screw fixation for treatment of the dens fracture under locational guidance is characterized by easy operation,minor trauma and satisfactory outcome.

Zuckerkandl tubercle is the extension of thyriod gland. For the past few years, more and more surgeons and anatomists have become interested in it. Clinically, surgeons usually rely on specified anatomical landmarks to avoid injuries to the recurrent laryngeal nerve, including the laryngeal branches and the superior parathyroid in thyroid surgery, such as the bottom corner of the thyroid cartilage, the inferior thyroid artery, lower pole of thyroid, tracheoesophageal groove, suspensory ligament of thyroid gland and so on. As the specified landmarks, they have several common characteristics:on the one hand, their location should be constant; on the other hand, they should be easy to observe and touch, meanwhile convenient and practical.As a new anatomical landmark, Zuckerkandl tubercle is in accordance with the above characteristics and prevalent in the population. Therefore, Zuckerkandl tubercle could become an important landmark to identify the the recurrent laryngeal nerve, including the laryngeal branches and the superior parathyroid in thyroid surgery. This essay aims to briefly analyze the role of Zuckerkandl tubercle in thyroid surgery.

Objective To discuss diagnosis and anterior surgical treatment of hyperextensian cervical spine injury combined with intervertebral disk injury. Methods A retrospective study was done on clinical data of 27 patients who suffered from hyperextension cervical spine injury combined with intervertebral disk injury to analyze their age distribution, clinical symptomes, X-ray and MRi manifesta-tions and perioperative intervertebral disk injury. All patients were treated with discectomy, strut bone grafting within vertebral bodies and internal fixation with titanium plate. The clinical outcomes were evalu-ated by using Frankel scale and ASIA motor score (AMS). Results Both MRI and X-ray detected following abnormal pathological changes in all patients: rupture of anterior longitudinal ligament, horizon-tal tear of disk, intervertebral disk hernia, compression and edema of spinal cord. The follow-up lasted for 9-32 months (average 17.5 months), which showed that all patients got improvement for 1-3 scales except that one patient with Frankel A had no improvement in neurological function. Compared with AMS on admission, both AMS at two months after surgery and at final follow-up was increased significantly, with recovery rate of AMS for 44.9% and 68.1%, respectively. There found no hardware related compli-cations such as implant loosening, defluxion or breakage. Bone fusion was found in all fixation segments. Conclusions MRI and X-ray are important examination means for hyperextension cervical spine injury combined with intervertebral disk injury. On a specified diagnosis, anterior surgical treatment should be done early and can get satisfactory recovery of spinal cord function.

AIM: To construct the prokaryotic expression system containing protein transduction domain (PTD) with heat shock protein 27 (HSP27) in order to prepare and purify the recombinant protein , and to verify whether the recombinant protein PTD-HSP27 has the ability to penetrate the human lens epithelial cell ( HLEC) membrane and the rabbit cornea.METHODS:The plasmid pKYB-PTD-HSPB1-6His was constructed by the technique of overlap extension PCR.The plasmid was transformed and PTD-HSP27 was purified through nickel affinity chromatography column and identi-fied by Western blotting.PTD-HSP27-6His was labeled with the fluorescein isothiocyanate (FITC).The penetrating ability of PTD-HSP27 into HLECs and rabbit cornea was tested .RESULTS:The recombinant PTD-HSP27 plasmid was success-fully cloned and effectively expressed .The correctness of the recombinant protein PTD-HSP27 was demonstrated .Fluores-cence microscopic examination showed that PTD-HSP27-FITC was internalized by HLECs .Fluorescent labeled PTD-HSP27 was then observed in the rabbit aqueous humor .CONCLUSION:The recombined gene PTD-HSPB1 was constructed by o-verlap extension PCR technique and the PTD-HSP27 fusion protein was prepared and purified by nickel affinity chromatog-raphy column.Using the technique of PTD-fusion protein, HSP27 was transduced into HLECs and passed through the cor-nea .

Objective To evaluate the clinical efficacy of a self-designed novel n-shaped aiming device in aiding the percutaneous cannulated screwing for treatment of odontoid fractures of Anderson-D' Alonzo type Ⅱ and shallow type Ⅲ.Methods A retrospective analysis was conducted of the 17 patients who had been treated at our department for odontoid fracture of Anderson-D' Alonzo type Ⅱ and shallow type Ⅲ between January 2009 and December 2013.They were 13 males and 4 females,with an average age of 37.5 years (from 16 to 61 years).All received surgical treatment after skull traction or jaw pillow traction preoperatively for reduction.Intraoperatively,percutaneous cannulated screwing via the anterior cervical approach was performed with the help of a self-designed novel n-shaped aiming device to fix the odontoid.Postoperatively,X-ray and CT scan were used to check the screw locations and evaluate the fracture healing.Results The 17 cases were followed up for 6 to 15 months (average,10.5 months).No intraoperative injury to the esophagus,artery,spinal cord or nerve root happened.The operating time ranged from 60 to 125 min.The bleeding was from 10 to 40 mL.Intraoperative observation revealed fine locations of the screws.Bony union was achieved in 16 cases 3 months postoperatively while separation and hardening of the fracture ends was found in one ease.Conclusion Our self-designed novel n-shaped aiming device can facilitate the percutaneous cannulated screwing for treatment of odontoid fractures of Anderson-D'Alonzo type Ⅱ and shallow type Ⅲ,given that it has advantages of simple manipulation,accurate insertion,minimal invasion,quick recovery and reliable efficacy.

Objective To explore the local application of immunosuppressant in improving the survival rate of the transplanted islet cells and systemic side effects.Methods The streptozocin of 200 ms/kg was injected into the abdominal cavity of the Wistar rats，the blood sugar was tested after 48，and 72 hours，and the rats with two consecutive measurements ≥20 mol/L were taken as the experimental animal model.The dose of pancreatic islet cells transplanted into the abdominal cavity was 8 000 IE,/kg，and that of cyclosporine dosage was 1.5 mg/(100 g·d).The pancreatic islet cells were divided into three groups：(1)systemic immunosuppressive agents through stomach lavage with the intraperitoneal injection of microencapsulated islet cells；(2)pure intraperitoneal injection of microencapsulated islet cells；(3)intraperitoneal injection microencapsulated activated carbon particles loaded with immunosuppressants，and mieroencapsulated islet cells.Changes of blood glucose and pathological in rats after transplantation were detected.Results The blood glucose of group 3 and group 1 showed no significant difference(P＞0.05)，as well as compared with group 2(P＞0.05).But the local application of immune agents could prolong the effective time of the islet cells and attenuate the fibrotic extent of the surrounding islets when compared with the control group,the C peptide level in applicating immunosuppressive agents group was significantly hisher in the immunosuppressive group than the pure transplantation group.ConclusionCompared with the systemic immune suppression via stomach lavage，local application of slow-release immunosuppressive agents showed the same effects of activated carbon particles，with a prolonged the effective time of islet cell and reduced topical side effects in the latter.