Background Recent research showed that stromal cell-derived factor-1 (SDF-1) plays critical role in mediating inflammation,metastasis of tumour and neovascularization of tumour.However,There is still no report about the research of the effects of SDF-1 on alkali-burn-induced corneal neovascularization (CNV).Objective The aim of this study attempts to investigate the role of SDF-1 and chemokine receptor CXCR4 in alkali-induced corneal neovascularization in mice.Methods Alkali-induced-corneal neovascularization animal models were constructed of in 15 eyes of 15 clean C57/BL mice by placing the filter paper with 1 mol/L NaOH to the central cornea for 10 seconds.The animals were sacrificed and specimens of cornea were obtained in 3,7,14 days after alkali burn of cornea.The expression of SDF-1/CXCR4 protein in the corneal tissues was detected by immunohistochemistry and Western blot,and expression of SDF-1/CXCR4 mRNA was detected by reverse transcription PCR (RT-PCR).Results SDF-1 was absent expressed and CXCR4 was faintly stained in normal cornea.The expression level of SDF-1 and CXCR4 was significantly enhanced in different time points after alkali burn in comparison with normal control mice by RT-PCR(P＜0.05),and the same trend was seen in the expression level of SDF-1 and CXCR4 proteins by Western blot (P＜0.05).The expression level of SDF-1 and CXCR4 in cornea peaked in the seventh day and began to decline in the fourteenth day but was still higher than normal level.Conclusion The study indicates that SDF-1/CXCR4 plays an important role in the formation of corneal new vessel in alkali-burn-induced animal model.

Objective The fluorescence-activated cell sorting (FACS) technique is a method for the identification and isolation of different cell populations.At present,the special surface marker for limbal stem cells has been not found yet.This study aimed to investigate the application of FACS technique in the research of rabbit limbal stem cells.MethodsCorneal limbal tissue was obtained from New Zealand white rabbits and cultured using the explant culture method in SHEM.Side population cells (SP cells) and non-SP cells were sorted from cultured rabbit limbal epithelium cells by FACS at a excitation wavelength 350 nm,and acquistion length 450 nm (blue light) and 675 nm (red light).The SP cells and non-SP cells were identified by detecting the expression of ABCG2 and K3/K12.The colony-forming efficiency of SP cells and non-SP cells were evaluated by the observation of cellular vitality with trypan blue staining.The percentage of colony formation was calculated as the colony number in various group/200×100%.ResultsIn 48-72 hours after primary culture,limbal epithelial cells migrated from the cultured tissue mass to form the mambrane-like structure and achieved 70%-80% confluence.The cells showed round,polygon and flattened shape.The proportion of SP cells in cultured limbal epithelial cells was 0.22%±0.09% with a colony-forming efficiency of 5.52±0.45% in SP cells and 0.78%±0.73% in non-SP cells,with a statistically significant difference between the two populations (t＝2.17,P＜0.01).After verapamil,an inhibitor of the expression of the ABCG2 protein,was added into the medium,the proportion of SP cells in the cultured limbal epithelial cells declined to 0.04%±0.006%.The SP cells presented a positive immunoresponse for ABCG2 and absence of immunoresponse for K3/K12,but a contradictory staining result was found in non-SP cells.ConclusionFACS can be applied in the research of limbal stem cells.

Objective:To investigate the effect of NS-398,a selective cyclooxygenase-2(COX-2) inhibitor,on the proliferation and invasion of human colon carcinoma cells in vitro,so as to determine the possibility of COX-2 as a new target for treatment of colon carcinoma.Methods: The expression of COX-2 in colorectal cancer cells(CW-2,COLO-320) was detected by RT-PCR and Western blotting.COLO-320 cell proliferation was measured by MTT after treatment with NS-398.Cell invasion ability was measured using migration and invasion chamber systems.Western blotting assay was used to examine the influence of NS-398 on MMP-2 expression.Results: Our results showed that CW-2,COLO-320 cells expressed COX-2 mRNA and protein.NS-398 inhibited the proliferation of COLO-320 cells in a time-and concentration-dependent manner.Invasion test showed that NS-398 inhibited the migration and invasion of COLO-320 cells.Western blotting revealed that NS-398 inhibited the expression of MMP-2 in COLO-320 cells.Conclusion: The selective COX-2 inhibitor NS-398 can inhibit COLO-320 cell proliferation and invasion,indicating COX-2 may serve as a new target for colon carcinoma treatment.

Objective To investigate the prevalence and correlation factors of cardiovascular damage in patients with diabetic nephropathy (DN) and non-diabetic nephropathy (NDN).Methods A total of 278 chronic kidney disease (CKD) patients admitted to the First Affiliated Hospital of Jinan University from January 2014 to May 2016 were enrolled,including 78 case of DN and 200 case of NDN.Patients had cardiac and carotid ultrasonography test by colour doppler ultrasonography,and their clinical and biochemical data were collected.Multiple linear regression analysis and multivariable logistic regression analysis were applied to study the correlation factors of cardiovascular damage in CKD patients.Results Mean age was 48.22 years in the 278-patient cohort,which included 178(64.03％) men.Compared with NDN group,DN patients had higher left atrial dimension,interventricular septal thickness,left ventricular end-diastolic dimension,left ventricular posterior wall thickness,left ventricular mass index (LVMI),carotid intima-media thickness (cIMT) and carotid plaques ratio.Their estimated glomerular filtration rate (eGFR) and the ratio between the peak speed of the early filling wave and that of the atrial contraction wave (E/A ratio) were however lower (all P ＜ 0.05).Prevalence of left ventricular hypertrophy (LVH),left ventricular relaxant function reduction and cIMT thickening in DN group were 67.95％,70.27％ and 57.14％,higher than those in NDN group (40.00％,42.31％ and 17.39％,respectively) (all P＜ 0.05).Along with the progress of CKD,LVMI and LVH proportion in patients with DN and NDN increased gradually.LVMI and LVH proportion in DN patients in CKD 1-2 phase and CKD 3-4 phase were higher than those in NDN patients (all P ＜ 0.05).In all CKD phases,cIMT and cIMT thickening proportion in DN group were higher than those in NDN group (all P ＜ 0.05).Just in CKD 1-2 phase,DN group had lower E/A ratio and higher proportion of left ventricular relaxant function reduction than NDN group (all P ＜ 0.05).After multiple linear regression analysis,gender,BMI,hemoglobin,eGFR and DN were related with LVMI;age,serum calcium and DN were related with E/A ratio;age and DN were related with cIMT (all P ＜ 0.05).In multivariate logistic regression,DN,hemoglobin and eGFR decrease were independently associated with LVH;age and BMI were independently associated with reduction of left ventricular relaxant function;age and DN were independently associated with cIMT thickening in all CKD patients (all P ＜ 0.05).Conclusions DN patients have more severe cardiovascular damage than NDN patients,and DN may be associated with LVMI,E/A ratio,cIMT,LVH and cIMT thickening in all CKD patients.

Objective To explore the effect on ultrasound localization method with radial artery cannulation in infants.Methods Sixty infants (1 month-1year old)undergoing elective surgical proce-dures required artery pressure,were randomly divided into two groups:ultrasound localization group (group B):the infants were used of portable B ultrasound probe in the wrist with two dimensional ul-trasound image to determine the body position of the radial artery;control group (group C):tradi-tional palpation determine the radial artery puncture position.One puncture success rate,total punc-ture success rate,the period of puncture time and incidence of complications were recorded;MAP, the radial artery diameter,the vertical distance from the ultrasound probe to the skin and the length from the puncture spot to the radial artery were also measured.Results The success rate of first puncture in group B was higher than that of group C (P <0.05),the times of puncture and procedure time were significantly less than those of group C (P <0.05),the incidence of hematoma was lower than that in group C (P <0.05).Conclusion The application of ultrasound-guided method for radial artery cannulation is safe and reliable with accurate localization,highly successful rate of puncture at first time,lower complication and shorter time for placement.

10.3969/j.issn.1008-9691.2013.03.007

Objective:The aim of this study is to investigate the effects of patients′ psychosocial background on the recurrence of aphthous ulcer (RAU). Methods: A total of 105 patients with RAU and 100 normal subjects of the control group were evaluated for psychosocial background. All subjects were required to complete the EPQ questionnaire, the SCL 90 questionnaire, and the enquired data were analyzed by using chi_square test.Results: Special personality and social factors in RAU patients might be reasons leading to the occurrence of this disease (P<0.01). Conclusion: Psychosocial disorder caused by stresses of social life events on the people with special personality could impact the occurrence of RAU.

BACKGROUND:The internal structures of the colagen-heparan sulfate scaffold and human nerve are very similar. OBJECTIVE: To explore thein vivo biocompatibility of colagen-heparin sulfate scaffold. METHODS:Forty pigs were randomly divided into two groups, 20 in each group: observation group and control group. Medulo-puncture needle was inserted 1.0 cm adjacent to the midline of anterior fontanele into the subarachnoid space, and then removed gradualy. Colagen-heparin sulfate scaffold was implanted into the observation group, and no treatment was given in the control group. Brain tissues were observed under transmission electron microscope, and cel apoptosis and Caspase-3 expression were detected at days 1, 3, 7, 14 and 30 after surgery. RESULTS AND CONCLUSION:Under the electron microscope, there were some damaged neurons in the observation group with the emergence of demyelination changes in the myelinated nerve fibers; positiveexpression of Caspase-3 protein was found at the junction between the brain tissue and scaffold as wel as within the scaffold, but no positive expression was found in the surrounding tissue. There was no cel apoptosis within 30 days after surgery except for individual apoptotic neurons both in the observation group and control group. The number of apoptotic cels in the observation group was higher than that in the control group at days 1, 3, 7, 14 days after surgery (P < 0.05), but there was no difference between the two groups at 30 days after surgery (P > 0.05). Caspase-3 protein expression was at a low state in the two groups, but the protein expression of Caspase-3 was higher in the observation group than the control group at days 3 and 7 after surgery (P < 0.05). These findings indicate that the colagen heparin sulfate scaffold has good biocompatibility in the porcine brain.

OBJECTIVE:To establish the method for simultaneous determination of 14 kinds of isoflavones in Pueraria radix. METHODS:LC-MS/MS was adopted to detect 14 kinds of isaflavones in 14 batches of P. radix(P. radix:PL-1 to PL-7,P. thomsonii:PT-1 to PT-7). The determination was performed on Extend C18 with mobile phase consisted of methanol-water(gradient elution)at the flow rate of 0.8 mL/min. The column temperature was set at 22℃. The sample size was 5μL. Ion source was ESI source. The detection mode was negative ion detection. Scanning mode was MRM with jet voltage of -4500 V;ion source temperature was set at 600 ℃, and atomization gas was 60 psi. The auxiliary gas was 60 psi,collision gas was 7 psi,air curtain gas was 30 psi. SIMCA 13.0 software wasused for cluster analysis of above batches. RESULTS:The linear range of 14 kinds of isoflavones ranged 10-1000 ng/mL(puerarin of 10-5000 ng/mL,r>0.9900). RSDs of precision,stability and reproducibility tests were all lower than 5％. The recoveries were 95％-105％(RSD were 0.8％-4.5％,n=6). The total content of isoflavones were different significantly between P. radix and P. thomsonii. The contents of isoflavones in P. radix from different origins were different significantly. Among 14 kinds of isoflavones, the content of puerarin was the highest. Results of cluster analysis showed that 14 batches of P. radix could be clustered into 4 categories,including PL-2 as Ⅰ category,PL-3 and PL-4 as Ⅱ category,PL-5,PL-6 and PL-7 as Ⅲ category,other as Ⅳcategory. CONCLUSIONS:The method is simple and reproducible. It can be used for content determination of 14 kinds of isoflavones in P. radix and quality control.

Objective:To summarize the best evidence for the application of massage in the management of meibomian gland dysfunction (MGD) -related dry eye.Methods:A systematic search was conducted in databases and websites including UpToDate, BMJ Best Clinical Practice, Joanna Briggs Institute Evidence-Based Healthcare Center database, National Guideline Clearinghouse, Medlive, PubMed, China National Knowledge Infrastructure, and WanFang data. The search covered literature on massage for MGD-related dry eye, including clinical decisions, best practices, evidence summaries, guidelines, expert consensus, systematic reviews, and randomized controlled trials. The search period extended to December 2023. Two researchers independently evaluated the quality of the literature and extracted and summarized the evidence.Results:A total of nine articles were included: three guidelines, four expert consensus papers, and two systematic reviews. Twenty-five pieces of best evidence were summarized from six aspects: massage practitioners, massage methods and indications, massage instruments and devices, massage duration and frequency, combination therapy, and the safety and efficacy of massage.Conclusions:This study summarizes the best evidence for applying massage in MGD-related dry eye. It is recommended that healthcare professionals apply this evidence in combination with clinical context and patient preferences.