Objective Explore the risk factors and prevention of HAP after surgical excision of Esophageal carcinoma in elderly Patients.Methods Two hundred and eight elderly patients with HAP after Surgical excision of Esophageal carcinoma from 2009 to 2012 were analyzed.Logistic regression analysis was used to find the related risk factors.Results The incidence of HAP was 25.9％.In several factors Logistic regression analysis,smoking,low lung function,postoperative non-epidural analgesia,injure of recurrent laryngeal nerve and fistula of anastomotic were the risk factors.Conclusions The preoperative evaluation,Intensive monitoring during the postoperative period and reducing the complication after operation can decrease the incidence of HAP.

Objective To monitor the release of amino acids of the whole retina during and after experimental glaucoma by increasing the intraocular pressure (IOP). Methods Experimental glaucoma was induced in one of the two eyes of rabbits by increasing IOP at 120 mm Hg for 45 min under infusion of saline in anterior chamber;then the pressure was released and the needle inserted into the anterior chamber was removed,this state was maintained for another 45 min.Every 15 min during the experiment 5 rabbits were killed and experimental eyes were enucleated.Aliquots (20 ?l) of the retinal extracts (see below) were mixed with ophthaldialdehyde reagent and analysed for amino acid content by the HPLC method of Wangwei, using a 150 mm?4.6 mm,5 ?m C18 column. Results A large increase in the release of glutamate,but not of the other three amino acids monitored,occurred during initial experimental ocular hypertension.It reached peak value of (111.73?17.46) 10 -5 mmol/g at 15 min of hypertension.15 min after release of intraocular pressure,again,immediately large and specific increase in the concentration of glutamate was reached to (102.96?51.91) 10 -5 mmol/g.In eyes subjected to paracentesis of anterior chamber,no difference was found between experimental eyes and controls. Conclusion These results suggest that glutamate is triggered by increasing the IOP,and it releases not only during the period of experimental ocular hypertension,but also afterwards.

ObjectiveTo analyze the protective effects of heat-shock response on the retinae of the rats after retinal ischemic reperfusion injury. MethodTwenty Wistar rats (20 eyes) were divided into 4 groups: intracameral perfusion group (group P), intracameral perfusion after quercetin injection group (group P+Q), intracameral perfusion after heat shock group (group P+H), and intracameral perfusion after quercetin injection and heat shock group (group P+Q+H). According to the standard program established by International Society for Clinical Visual Electrophysiology, we recorded the results of the dark-adapted electroretinogram (D-ERG),oscillatory potentials (OPs),and light-adapted ERG (L-ERG) of the rats with intraocular hypertension after induced by heat shock response. The expressions of HSP 70 of the rats in all groups were observed by Western blotting.ResultsThe expression of HSP 70 of the rats in group P+H was the highest in all groups, but the expressions of HSP70 in group P+Q and P+Q+H were inhibited significantly. The amplitudes of a and b wave of ERG and O 2 wave of OPs decreased, and the delitescence of them were delayed significantly in rats after intracameral perfusion. The amplitude of b wave of D-ERG and O 2 wave of OPs in group P+H were higher than which in group P. Zero hour after perfusion, the amplitudes of all waves in group P+H increased significantly (P

Objective To investigate the influence of live combined bifidobacterium, lactobacillus and enterococcus powder assisted with nasal jejunum nutrition on laboratory index, complication rate and economical efficiency in patients with severe acute pancreatitis (SAP). Methods Fifty patients with SAP were divided into treatment group and control group by random draw method with 25 cases each. The patients of 2 groups received conventional therapy of SAP and nasal jejunum nutrition, and the patients in treatment group were treated with the above treatment combined with live combined bifidobacterium, lactobacillus and enterococcus powder. The laboratory indexes, gastrointestinal function score, complications, hospitalization time and hospitalization expenses were compared between 2 groups. Results The white blood cell, amylase, lipase, C reactive protein, interleukin (IL)-8, tumor necrosis factor (TNF)-α, lactate dehydrogenase (LDH) and gastrointestinal function score after treatment in treatment group were significantly lower than those in control group:(5.9 ± 2.1) × 109/L vs. (8.4 ± 3.1) × 109/L, (210.4 ± 47.6) U/L vs. (271.9 ± 82.2) U/L, (205.2 ± 22.3) U/L vs. (249.3 ± 34.7) U/L, (14.7 ± 0.4) mg/L vs. (35.1 ± 0.8) mg/L, (16.0 ± 4.8)μg/L vs. (36.5 ± 12.9)μg/L, (21.7 ± 5.6) ng/L vs. (43.4 ± 9.5) ng/L, (212.5 ± 95.4) U/L vs. (284.0 ± 124.6) U/L and (0.81 ± 0.24) scores vs. (1.37 ± 0.36) scores, and the total incidence of complications, hospitalization time and hospitalization expenses were significantly lower than that in control group: 12.0%(3/25) vs. 64.0%(16/25), (18.72 ± 1.90) d vs. (21.13 ± 2.35) d and (4.48 ± 0.55) × 104 yuan vs. (4.73 ± 0.78) × 104 yuan. There were statistical differences (P<0.05). Conclusions Live combined bifidobacterium, lactobacillus and enterococcus powder assisted with nasal jejunum nutrition in the treatment of SAP patients can efficiently improve the laboratory indexes, promote gastrointestinal function recovery, decrease the risk of complications and reduce the economic burden.

Objective To investigate the expressions of Hedgehog sisnaling pathway genes in hepatocellular carcinoma tissues(HCC),and the effect of specific Hedgehog pathway inhibitor(KAADcyclopamine)on the growth of HCC cells and the expressions of Hedgehog genes. Methods The expression of Hedgehog signaling pathway components(Ihh,Ptch,Smo and Gli)was investigated in 14 HCC tissue slices,4 HCC cell lines and a normal hepatic cell line by using immunochemistry.The expression of Ihh,Ptch,Smo and Gli proteins was investigated in 9 HCC tissue specimens and 6 normal hepatic tissue specimens by using Western blotting.The expression of Ihh、Ptch、Smo、Gli and Hip genes was investigated by RT-PCR.Results The positive ratio of Gli,Ptch,Ihh and Smo were 42.9%,71.4%,71.4% and 85.7% in 14 HCC tissue slices,respectively.The expressions of Gli protein and Gii gene were up regulated while the expression of Hip gene was down regulated in HCC specimens compared with normal hepatic tissue specimens.Hedgehog signaling pathways in HCC cell lines HepG2,Bel-7402 and QGY-7701 were activated;KAAD-cyclopamine,a specific inhibitor of the Hedgehog signaling pathway,down regulated cell growth and the expressions of Ptch and Gli genes in the 3 HCC cell lines(Ptch gene:tHepG2=3.78,tBel-7402=9.03,tQGY-7701=5.63;Gli gene:tHepG2=9.61,tBel-7402=4.15,tQGY-7701=20.30,P＜0.05 in each group).The expression of Hip gene was up regulated in QGY-7701 after treated with KAAD-cyclopamine(t=4.70,P＜0.05).Conclusion The expression of main Hedgehog signaling pathway components were detected in HCC,KAAD-cyclopamine specifically inhibited the Hedgehog signaling pathway.

Background Anti-scarring is a focus following glaucoma filtering surgery.It has been verified that expression of heat shock protein 47 (HSP47) is always correlated with that of various types of collagens and closely related with the collagen-related diseases including fibrosis in various organs.However,the relationship between the expression of HSP47 and fibrosis of filtering bleb is unclear.Ohjective Present study was to investigate the dynamic expression of HSP47 in subconjunctival filtering bleb after anti-glaucomatous filtering surgery.Methods Filtering surgery was performed on the right eyes of 10 SPF male Spargue-Dawley(SD) rats.Then the operative eyes were randomized into the postoperative 7-day group and 14-day group according to the randomized number table,and the left eyes of the rats served as the normal control group.The shape of filtering blebs and the ocular anterior segment response were examined under microscope daily.The animals were sacrificed by excessive anesthesia on day 7 and 14 after operation,respectively,and the specimens were prepared of the eyes.Regular histopathological examination was performed to observe the collagen fibrosis change,and immunohistochemistry was carried out to assay the dynamic expression of HSP47 depended on the fibrosis.Percentage of positive cells for HSP47 were calculated.The use and care of the experimental animals complied with the Regulation for the Administration of Affair Concerning the Experimental Animals by State Science and Technology Committee.Results Diffuse subconjunctival blebs were seen postoperatively in t0 eyes after surgery with mild inflammatory response in ocular anterior segment.Hematoxylin-eosin staining showed that compared with the specimens of the normal control group,fibroblasts obviously proliferated and collagenous fibers increased in the specimens of operative group.The HSP47 protein was less expressed in cytoplasm of many fibroblasts and less vascular endothelial cells in the normal rat specimens.The expressing intensity of HSP47 protein was enhanced.However,the expression intensity was weaker in the postoperative 14-day group in comparison with the 7-day group.The percentage of positive cells for HSP47 in the filtering blebs 7 days after operation was (56.40±1.12)％,that in the normal control group was (13.70±0.74)％,with a significant difference between them (P=0.000).In the postoperative 14-day group,the percentage of positive cells for HSP47 reduced from (56.40± 1.12) ％ to (23.90±0.76) ％ (P =0.000),but it was still higher than that of the normal control group (P =0.000).Conclusions Expression intensity of HSP47 alters as the activity of fibroblasts in filtering bleb,suggesting that HSP47 plays a role in collagen-fibrosis following filtering surgery in SD rats.

Background Misfolding of Myocilin protein plays an important role in the pathogenesis of primary open angle glaucoma (POAG).GRP78 can transfer the misfolded proteins out endocytoplasmic reticulum and keep the protein synthesis function of endoplasmic reticulum.However the relationship between GRP78 and Myocilin in the pathogenesis of POAG has not been elucidated.Objective This study was to investigate and analyze the coexpressions of GRP78 and Myolicin in trabecular meshwork cells (TMCs) of POAG.Methods The trabecular meshwork tissues were obtained during the surgery of POAG and healthy donor for the isolated and in vitro culture of TMCs,and the morphology of the cells was compared between POAG-derived TMCs and donor-derived TMCs.The expression of specific factors for TMCs were detected by immunochemistry.The cells were divided into normal control group,tunicamycin (Tm)-treated group and staurosporine (STS)-treated group,and the cells were cultured by regular medium,the 1 μ mol/L Tm medium or 0.1 μmol/L STS medium for 24 hours respectively.The co-expression of GRP78 and Myocilin in the cells were detected using immunofluorescence assay.This study was approved by Ethics Committee of Xi'an No.4 Hospital,and written informed consent was obtained from each patient before surgery.Results Primarily cultured cells showed the fiat star-like and irregular appearance with 3-5 processes,round nuclei,abundant cytoplasm and black engulf particles.The 3-7 generations of POAG-derived cells grew well,showing positive expressions of laminin (LN),fibronectin (FN),neuronal specific enolase (NSE) and Vimentin.Immunofluorescence assay showed positive expressions of GRP78 and Myocilin in the cells of the Tm group,STS group and normal control group,with the reddish and green fluorescence separately.Incomplete co-expression of GRP78 and Myocilin was seen in donor-derived TMCs,and complete co-expression of GRP78 and Myocilin was found in the POAG-derived TMCs in the normal control group.Complete co-expression of GRP78 and Myocilin was exhibited in both donor-and POAG-derived TMCs in the Tm group and STS group.In addition,accumulation of GRP78 protein around nucleus was seen in both donor-and POAG-derived TMCs in the Tm group.Conclusions GRP78 and Myocilin proteins are completely co-expressed in POAG-derived TMCs,inferring that GRP78 and Myocilin play an interaction role in the pathogenesis and development of POAG.GRP78 may play a cytoprotective role against the apoptosis of TMCs caused by endoplasmic reticulum stress.

Background Ocular alkali burns leads to corneal ulcer and angiogenesis and even corneal opacity.There is still no ideal treatment method.Studies showed that mesenchymal stem cells (MSCs) can repair corneal wound in vivo,but the specific mechanism is still not clear.Objective This study aimed to observe the histopathological change after the early transplatation of bone marrow MSCs (BMSCs) for corneal alkali burn model in rabbits and explore the anti-inflammatory effects of MSCs after corneal alkali burn.Methods Bone marrow of 4 ml was collected from 2-3 month-old Japanese rabbit.BMSCs were isolated and cultured from the bone marrow of rabbits,and the third generation of cells were used in this study.Cultured cells were identified by morphology and the expressions of surface markers.Corneal alkali burn models were extablished in the right eyes of 24 rabbits by attaching the filter paper with 0.1％ NaOH at the central cornea for 30 seconds,and then the models were randomized into 2 groups.BMSCs suspension of 300 μl (concentration 5×l06/μl) was subconjunctivally injected 1 hour after modeling in the BMSCs group,and equal volume of PBS was used in the same way in the PBS group.Corneal opacification was scored under the slim lamp microscope in 3,14 and 28 days after injection.The polymorphonuclear neutrophils (PMNs) were counted by histopathological examination,and the expression of matrix metalloproteinase-2 (MMP-2) in the corneal tissue was evaluated by immunochemistry in various time points.The use and care of the rabbits followed the statement of ARVO.Results The rabbit BMSCs were plastic-adherent cells that exhibited a fibroblastlike shape.Cultrued cells highly expressed surface adhesion molecular markers CD29 and CD90 (99.18％ and 97.94％) and lowly expressed hematopoietic cell markers CD34 and CD31 (0.74％ and 0.15％).Opacification of cornea,defect of corneal epithelium,stromal edema and neovascularization appeared after modeling.In 14 days and 28 days after modeling,the opacification scores in the BMSCs group were 2.37±0.52 and 2.25±0.50,which were significantly lower than 3.00±0.53 and 3.25 ±0.50 in the PBS group (t =2.376,2.828,both at P＜0.05).After subconjunctival injection,the number of PMNs was (34.17 ±1.85) /12 fields and (25.64 ±3.86)/12 fields in the BMSCs group,showing significant decrease in comparison with (42.70 ±1.54) /12 fields and (32.67 ±1.42)/12 fields in the PBS group (t=10.021,4.832,both at P=0.000).The expression levels of MMP-2 (A value) in cornea were 0.388±0.016 and 0.384±0.006 in the BMSCs group,with considerable decreases in comparison with 0.438± 0.006 and 0.412± 0.005 in the PBS group (t=10.205,13.514,both at P=0.000).Conclusions Early transplantation of BMSCs can arrest the occurrance of corneal ulcer by suppressing the infiltration of PMNs,alleviateing the inflammation reaction,downregulating the expression of MMP-2 in cornea and inhibiting the degradation of stromal collagen fibers.

Objective To evaluate the effects and underlying mechanisms of crocin on glutamate-induced apoptosis of retinal ganglion cells (RGCs) by affecting extracellular calcium influx.Methods Primary rat retinal ganglion cells were isolated and stimulated with glutamate at concentrations of 0.1 mmol/L and 1 mmol/L for 24 h or 48 h,respectively,to establish apoptosis model of RGCs.Afterwards,crocin of different doses (0.1,1.0 and 3.0 μmol/L) was used to treat the glutamate-induced RGCs for 12 h;then cell apoptosis was detected by Annexin V-FITC/PI staining.The intracellular calcium concentration was determined by FIuo-3/AM fluorescent labeling.Western blot was used to examine the effect of crocin on Ca2+-mediated apoptotic signal molecules calpain and CaMKII.The mitochondrial membrane potential was detected by JC-1 staining and mitochondrial apoptosis-related signaling molecules Caspase-3,Caspase-9 and Bcl-2/Bax were evaluated by Western blot,respectively.Results In comparison with the untreated controls,the cell apoptosis of RGCs exposed to 0.1 mmol/L of glutamate for 24 h did not significantly change (P＞ 0.05).However,apoptosis rate of the cells reached (43.050 ± 2.616) ％ when the stimulation time lasted for 48 h and showed a significant increase (P＜0.01).Treatment with higher-dose glutamate (1 mmol/L) significantly increased apoptosis of RGCs at 24 h (46.450±1.061)％ and 48 h (45.500±3.253)％ compared with the controls (P＜0.01).RGCs were induced by 1 mmol/L of glutamate for 12 h,followed by the treatment with crocin at concentrations of 0.1,1.0 and 3.0 μmol/L,respectively.Each dose of crocin could significantly inhibit cell apoptosis in the dose-dependent manner (P＜0.01).In addition,crocin at 1.0 μmol/L blocked glutamate-induced extracellular calcium influx,inhibited the expression of calcium-dependent proteins Calpainl and CaMK Ⅱ.Moreover,crocin at the dose of 1.0 μmol/L also increased mitochondrial membrane potential,suppressed the expressions of Caspase-3 and Caspase-9,and elevated Bcl-2/Bax ratio.Cornclusion Crocin inhibits glutamate-induced apoptosis of retinal ganglion cells through suppressing extracellular calcium influx,thereby blocking calcium-dependent and mitochondria-dependent apoptosis signaling pathways.

Objective To explore the application value of Carbon Nanoparticles followed by microscale Methylthioninum Chloride in sentinel lymph node biopsy for breast cancer.Methods Two hundred and eight breast cancer patients were selected in the First Affiliated Hospital of Zhengzhou University during recent two years who performed sentinel lymph node biopsy.They were randomly divided into five groups according to different lymphatic tracer injected:Sequential group (A group,128 patients),Carbon Nanoparticles group (B group,44 patients) and Methylthioninum Chloride group (C group,36 patients).According to different Methylthioninum Chloride doses,sequential groups were randomly divided into low dose group (A1 group,48 patients),middle dose group (A2 group,42 patients),high dose group (A3 group,38 patients),If sentinel lymph nodes were negative no longer performed axillary lymph nodes dissection.Receive the five groups statistics of detection rate,the condition of lymphatic tracer,the incidence of local skin irritation and so on.Results (1) The detection rate of A,B and C three groups had no obvious difference.The incidence of lymphatic tracer dyed not ideal in B group was higher than A and B two groups,the incidence of local skin stimulation in C group was higher than A and B two groups.(2) A1,A2,A3,three groups had no difference of the detection rate and lymphatic tracer,but the incidence of local skin stimulation in A3 group is higher than A1,A2 two groups.3.5 groups of SLN detection rate had no relationship with patients' age,BMI,tumor size,tumor location,tumor molecular classification factors (P ＞ 0.05).Conclusions Carbon Nanoparticles followed by microscale Methyhhioninum Chloride detection rate was similar with two separate application,but can reduce the lymphatic vessels not developing situation when used Carbon Nanoparticles alone and reduce the local irritation caused by large dose of Methylthioninum Chloride,so it can be a good choice of SLNB in clinical.