Background Researches have demonstrated that dexamethasone (Dex) can induce the changes of the function and structure of trabecular meshwork cells,and latrunculin A (Lat A) can enhance the outflow of aqueous humour and therefore low the intraocular pressure.Objective The aim of the present study is to investigate the effects of Dex and Lat A on the expression of protein in human trabecular meshwork cells.Methods Human trabecular meshwork cells were primarily cultured in DMEM using expand culture method and the fifth generation of cells were used to this experiment.Dex and/or Lat A were added in medium as 10~(-6)mol/L Dex group(Dex treating for 24 hours),Dex+Lat A group(10~(-6)mol/L Dex+2mmol/L Lat A for 24 hours),Lat A group(2mmol/L Lat A for 24 hours) and DMEM culture group.Two dimensional gel electrophoresis(2 DE) was used to compare the protein expressions among these four groups.Subsequently protein spots with different intensity were selected for mass spectrometry analysis.Results Four gel patterns of two dimensional gel electrophoresis of human trabecular meshwork cells from Dex,Dex+Lat A,Lat A and control groups were obtained.A good isolated result for majority of proteins in human trabecular meshwork cells was found in all of the four groups.An obvious expression difference of proteins in human trabecular meshwork cells was seen among the different culture conditions.Twenty four kinds proteins were identified by GDPiMALDI TOF MS,including cytoskeleton related proteins,heat shock proteins,redox related proteins,and proteins participating in carbohydrate metabolism.The expressions of aldehyde dehydrogenase(ADLH)and Rab were increased in Lat A group and decreased in Dex group,but HSP27 and hCRMP2 showed the contrary outcome.Conclusion This study construct the pattern of protein expression in human trabecular meshwork cells by using 2 DE.Dex and Lat A impact the protein expressions in human trabecular meshwork cells.

BackgroundExcessive conjunctival scar formation is a main cause of filtering surgical failure in glaucoma.It has been reported that the failure rate of filtering surgery is a tough problem in the research of glaucoma.Research showed that tetrandrine (Tet) suppress the proliferation of cultured human fibroblasts of Tenons capsule (TCFs) in vitro,but its possible mechanism is still unclear up to now.ObjectiveThe aim of this study is to investigate the inducement effects of Tet on the apoptosis of fibroblasts of Tenons capsule and mechanism.MethodsThe Tenon's capsule was obtained from donor eyes of Zhongshan Eye Center Eye Bank for further use.Human Tenon's capsule fibroblasts were cultured by explant culture method in mixed medium in vitro and the third to seventh generations of cells were collected for the experiment.The subcultured cells were identified by morphology observation and Vimentin staining.The apoptosis of cultured fibroblasts in Tet-treated group and control group was studied by using TUNEL (TdT- mediated dUTP nick end labling,TUNEL),flow cytometry (FCM) and transmission electron microscope.The cell number in different cellular cycle was calculated in Ted-treated group and control group.Results The cultured cells reached confluence in two weeks after cultured and presented the spindle or triangle shape with the radial-like or vortexin-like arrangement .The cells showed the positive staining for Vimentin.Apoptosis changes of cultured cell and apoptosis bodies were seen under the transmission electron microscope.Apoptotic cell nuclei were observed in Tet group by TUNEL.FCM result showed that the cells at G_0/G_1 phase decreased by 22.2%,and the cells at S and G_2/M phases increased by 20.53% and 1.6% in Ted-treated group.Significant differences in the numbers of cells in different cellular cycles and cell numbers of apoptosis were found between Tet-treated group and control group(P=0.000).ConclusionTet can inhibit the proliferation of human TCFs by inducing apoptosis in vitro.

ObjectiveTo investigate the in vivo biological performance of 5 porous bioceramic scaffolds,which were bioglass,β-tricalcium phosphate (β-TCP),hydroxyapatite (HA),β-calcium silicate (β-CS) and α-CS,implanted in rabbit dorsal muscle.MethodsThe 5 porous bioceramic scaffolds were fabricated by adding pore-forming materials and sintering,and then were investigated by X-ray diffraction,porosity mensuration and biomechanics test.The scaffolds were implanted into rabbit dorsal muscle for 4,8,12,16 weeks,respectively.The samples were analyzed by X-ray,Micro-CT,histological analysis,scanning electron microscope (SEM) and energy dispersive spectrometer (EDS).The expression of bone morphogenetic protein(BMP-2) and BMP-7 in the muscle in touch with bioceramic scaffolds were also investigated by polymerase chain reaction(PCR).ResultsThe characteristic analysis of 5 scaffolds showed that the sequence of compressive strength was bioglass＞α-CS＞β-CS＞β-TCP＞HA,the sequence of elasticity modulus was α-CS＜β-TCP＜HA＜β-CS＜bioglass.It was confirmed by X-ray,Micro-CT and histological analysis that the sequence of biodegradability was β-CS＞α-CS＞β-TCP＞bioglass＞HA.The histological observation showed no new bone formation in five scaffolds.A Ca-P layer was formed in the surface of bioglass,α-CS and β-CS,which suggested their in vivo bioactivity.After 16 weeks,the expression of BMP-2 and BMP-7 was found only in β-CS.Conclusion The porous calcium silicate scaffold,which was promising for bone tissue engineering,was with good in vivo bioactivity and biodegradability,without in vivo osteoinductivity.

Background Blepharitis caused by Demodex infestation is very common in clinical practice.There are various methods mentioned in the study of Demodex infestation in China,but a unified introduction and evaluation of the operating procedures is lacked.A quick and accurate clinical diagnostic method for Demodex infestation needs to be further studied.Objective This study aimed to establish operation procedures for the clinical examination of eyelid Demodex infestation,which were applied to evaluate the conditions of eyelid Demodex infestation in ocular patients with discomfort.Methods One thousand and fifty-two patients with eye dryness,eye itchiness or other symptoms were selected for slit lamp examination and photographing of the eyelid margin area.Three eyelashes with associated scurf from each superior eyelid were plucked out for examination of Demodex under the microscope.Positive findings included observation of Demodex mites or eggs.Their amounts were recorded individually for all eyelash samples.Results A procedure for observing,recording and reporting eyelid Demodex infestations in patients was successfully established.By using this procedure,1 052 patients were investigated for the examination of Demodex infestations.Demodex mites or eggs were found in 582 cases (55.3％).The positive rate of Demodex infestation increased with age,and the population over 60 years group had the highest positive rate,showing a significant difference among the different age groups (x2=10.547,P=0.001).There was no significant difference in positive rate between male patients and female patients (P =0.352).The test turnaround time (TAT) for one examination was (11.4±5.2) seconds.Conclusions The operational procedure for examining the palpebral margin Demodex infestation by the slit-lamp,optical microscope,photographing and laboratory reports is established.It is simple and quick in the appliation for the clinical diagnosis of eyelid Demodex infestation.

Objective:To investigate the relationship between the expression of serum exsomal miRNA-155-5p (miR-155-5p) and prognosis in patients with esophageal squamous cell carcinoma (ESCC).Methods:A total of 336 samples from ESCC patients in Fujian Provincial Cancer Hospital from October 2014 to December 2015 were collected. The relative expression levels of serum exsomal miR-155-5p were detected by using real-time fluorescent quantitative polymerase chain reaction (qRT-PCR). Cut-off value of the expression levels of serum exsomal miR-155-5p was determined by using X-tile software. Based on the optimal cut-off value, patients were divided into miR-155-5p low expression group and miR-155-5p high expression group. The survival curve was drawn by using Kaplan-Meier method and Cox proportional hazards regression model was used to make survival analysis.Results:The cut-off value of serum exsomal miR-155-5p expression level was 2.340. According to the cut-off value, patients were divided into miR-155-5p low expression group (<2.340) of 51 cases and miR-155-5p high expression group (≥2.340) of 285 cases. There were no statistical differences in age ( χ2 = 0.020, P = 0.887), gender ( χ2 = 0.283, P = 0.595), tumor location ( χ2 = 0.063, P = 0.977), differentiation grade ( P = 0.474), clinical staging ( χ2 = 3.996, P = 0.136) and surgery treatment ( χ2 = 0.941, P = 0.332) of patients in both groups. ESCC patients in serum exsomal miR-155-5p high expression had a higher risk of death compared with patients in miR-155-5p low expression group ( HR = 1.763, 95% CI 1.049-2.961, P = 0.032). Conclusion:The high expression level of serum exsomal miR-155-5p is associated with poor prognosis in ESCC patients and it could be used as a prognostic new marker in ESCC patients.

OBJECTIVE:To observe clinical efficacy and safety of Xuebijing injection combined with octreotide and ulinastatin in the treatment of acute severe pancreatitis.METHODS:A total of 150 patients with acute severe pancreatitis admitted to emergency department of our hospital during Jan.2012-Jan.2016 were divided into control group,drug control group and observation group according to therapy method,with 50 cases in each group.All patients were treated with fasting,gastrointestinal decompression time,antibiotics,blood purification and other conventional treatment.Control group additionally received Cctreotide acetate injection 0.1 mg intravenously,tid,on the basis of conventional treatment.Drug control group additionally received Ulinastatin for injection 100 000 U added into 10％ Glucose injection 250 mL,ivgtt,bid,on the basis of control group.Observation group additionally received Xuebijing injection 100 mL added into 10％ Glucose injection 100 mL,ivgtt,bid,on the basis of drug control group.Three groups were treated for 10 days.The clinical indexes as fotal response rates,gastrointestinal decompression time,abdominal pain relief time,length of stay were observed in 3 groups.Related serum indexes (AMY,WBC,IL-6,CRP,TNF-α) before and after treatment and the occurrence of compliance during treatment were compared among 3 groups.RESULTS:The total response rate of observation group was 90.0％,which was significantly higher than 72.0％ of drug control group and 52.0％ of control group,with statistical significance (P＜0.05).The gastrointestinal decompression time,abdominal pain relief time and length of stay in observation group were significantly shorter than drug control group and control group,with statistical significance (P＜0.05).Before treatment,there was no statistical significance in the serum levels of AMY,WBC,IL-6,CRP or TNF-α among 3 groups (P＞0.05).After treatment,above serum indexes of 3 groups were decreased significantly,and observation group was significantly lower than the drug control group and control group,with statistical significance (P＜0.05).The incidence of ARDS,shock and acute renal failure in observation group were significantly lower than drug control group and control group,with statistical significance (P＜0.05).There was no statistical significance in the incidence of sepsis,abdominal abscess or MODS among 3 groups (P＞0.05).CONCLUSIONS:Xuebijing injection combined with octreotide and ulinastatin show significant therapeutic efficacy for acute severe pancreatitis,can effectively control inflammation progress,improve clinical symptoms and promote disease recovery with good safety.

Objective To investigate the internal and external factors of scientific and technological innovations in pro-vincial maternal and child health care institutions and propose effective strategies for facilitating the innovations.Methods The PEST-SWOT model was used to analyze the internal strengths and weaknesses,external opportunities,and threats of the scientific and technological innovations in a provincial maternal and child health care institution from the perspectives of political,econom-ic,social,and technical environment.Results The institution has advantages and opportunities in policy support,scientific re-search management and investment,and disciplinary characteristics.However,it faces some threats and has disadvantages in tal-ent team,information-oriented level,and industrial competition.Conclusion The development of scientific and technological in-novation in provincial maternal and child health care institutions is affected by internal and external factors.It is necessary to firmly seize opportunities and comprehensively promote scientific and technological innovations from the aspects of medical treat-ment,teaching and research collaboration,discipline layout,talent team,and information-oriented construction.

OBJECTIVETo investigate the effect of tetrandrine (Tet) on the expression of bax, bcl-2, and transforming growth factor-β2 (TGF-β2) mRNA in cultured human fibroblasts of Tenon's capsules (TCFS) and explore its possible mechanism.

METHODSThe third passage of TCFS cultured in vitro were exposed to 1×10(-5) mol/L Tet for 24 h, and real-time fluorescence quantitative PCR was used to detect the changes in the expressions of bax, bcl-2, and TGF-β2 mRNA.

RESULTSThe expression level of bax mRNA was obviously higher, while bcl-2 and TGF-β2 mRNA levels were significantly lower in Tet-treated TCFS than those in the control cells (P<0.05).

CONCLUSIONTet can inhibit the proliferation of TCFS possibly by reducing the expressions of bcl-2 and TGF-β2 mRNA, enhancing the expression of bax mRNA and inducing cell apoptosis, suggesting its potential in preventing fibrous scar formation after glaucoma filtration surgery.

Apoptosis ; Benzylisoquinolines ; pharmacology ; Cell Proliferation ; drug effects ; Cells, Cultured ; Cicatrix ; metabolism ; prevention & control ; Fibroblasts ; cytology ; drug effects ; metabolism ; Humans ; Proto-Oncogene Proteins c-bcl-2 ; genetics ; metabolism ; RNA, Messenger ; genetics ; metabolism ; Tenon Capsule ; cytology ; Transforming Growth Factor beta2 ; genetics ; metabolism ; bcl-2-Associated X Protein ; genetics ; metabolism

Laboratory animals are an important part of life sciences and medical researches, as well an important support for the science and technology innovation in our country. Laboratory animal science is of great significance to the protection of human health,food safety and biological safety. Laboratory animals are indispensable in the development of food safety,drugs,vaccines and biological products and the studies of human disease pathogenesis. In order to adapt to the requirements for overall development of the laboratory animal industry in China, our institute has independently developed the Network Training System for Laboratory Animal Managers. This system is an online education and training platform which integrates the practical operation and theoretical knowledge of laboratory animals,including seven knowledge modules such as animal welfare,animal breeding,animal surgery and so on. The training subjects of the system include managers, experiment operators, laboratory animal doctors and breeders, aimed at accelerating the personnel training and team building of laboratory animal sciences,and promoting the transformation and development of personnel training in laboratory animal industry.

Objective:To investigate the therapeutic efficacy and potential mechanisms of integrin α vβ 3-targeted radionuclide therapy (TRT) in combination with anti-programmed cell death protein ligand 1 (PD-L1) immunotherapy. Methods:Integrin α vβ 3-targeted molecule Arg-Gly-Asp (RGD) was conjugated with Evans blue (EB) and then labeled with 177Lu to obtain 177Lu-EB-RGD. The radioactivity and radiochemical purity were determined. MicroSPECT imaging, biodistribution, and in vivo therapeutic efficacy were subsequently performed in MC38 murine colon cancer models. Volume of tumor and body mass of mice were observed to assess the therapeutic efficacy and safety ( n=9 in each group). Flow cytometry was used to evaluate therapy response of saline-treated (control, group A), 18.5 MBq 177Lu-EB-RGD-treated (group B), 10 mg/kg PD-L1 antibody-treated (group C), TRT combined with immunotherapy-treated (group D, 18.5 MBq 177Lu-EB-RGD and 10 mg/kg PD-L1 antibody) mice and alterations in tumor microenvironment (PD-L1 + immune cells, CD8 + T cells and regulatory T cells). Independent-sample t test and repeated measures analysis of variance were used for data analysis. Results:The radioactivity of 177Lu-EB-RGD was (55.85±14.00) GBq/μmol. SPECT imaging clearly visualized the MC38 tumors in mice models with high uptake and long retention time, the tumor/muscle ratio reached 14.87±0.88 at 24 h postinjection, while less uptake and retention in normal tissues. Tumor uptake of 177Lu-EB-RGD was significantly higher than that of 177Lu-RGD 4 h post-injection ((12.00±1.60) vs (3.69±0.37) %ID/g; t=8.63, P<0.01). The efficacy results between each treatment group was significantly different ( F=7.32, P=0.03) at day 6 post-treatment. The combination therapy showed the most outstanding anti-tumor efficacy with 7/9 mice showed complete response. Flow cytometry results showed that TRT up-regulated the PD-L1 expression significantly, namely, PD-L1 + immune cells in group B and group A were significantly different (CD45 + /PD-L1: 2.34% vs 0.95%, CD11b + /PD-L1: 2.41% vs 0.66%; t values: 11.17 and 8.70, both P<0.01); immunotherapy and combination therapy dramatically stimulated the infiltration of CD8 + T cells (2.07% vs 0.26%, 2.71% vs 0.26%; t values: 4.10 and 6.03, both P<0.05). Conclusion:TRT in combination with immunotherapy synergistically enhance anti-tumor efficacy, which is expected to play a role in the treatment of patients with advanced tumor where TRT can be applied.