Background Mesenchymal stem cells (MSCs) have been applied in basic and clinical researches of organ transplantation.Our previous study showed that intravenous injection of MSCs prolonged corneal allograft survival in rat.However,the effect of local administration of MSCs on corneal allograft rejection remains unclear.Objective The aim of this study was to investigate the effect of subconjunctival injection of MSCs on corneal allograft rejection in rat model of keratoplasty.Methods MSCs were isolated and cultured from femur and tibia bone marrow of clean Wistar rats,and then the cells were identified by induced differentiation of osteoblast and adipocyte.The third generation of MSCs was used in subsequent experiment.Allogenic penetrating keratoplasty was performed with the bilateral corneas of 20 Wistar rats as donor grafts and the right eyes of 40 Lewis rats as recipients.PBS 0.1 ml containing 2 × 106 MSCs and 0.1 ml PBS only was subconjunctivally injected immediately and postoperative 3 days respectively in randomized two groups,and another 6 normal Lewis rats served as the normal control group.Corneal rejection response was evaluated under the slit lamp after surgery based corneal opacity,edema and neovascularization,and the grafts were scored according to the criteria of Larkin.The corneal samples were extracted from 12 rats of the PBS control group and the MSCs group separately 10 days after surgery.The relative expressions of Th1 cytokines (interferon-γ [IFN-γ] mRNA and interleukin-2 [IL-2] mRNA) and Th2 cytokines (IL-4 mRNA and IL-10 mRNA) were detected by real-time quantitative PCR.Protein levels of IL-4 and IL-10 proteins in the corneas were assayed by ELISA.All experimental protocols involving rats were approved by the laboratory animal care and use committee of the Tianjin Medical University and treated with the ARVO statement for the use of animals in ophthalmic and vision research.Results The cells grew well with the orange stain for alizarin red in differentiated the osteoblasts and red stain for Oil red O in differentiated adipocytes.The survival time of corneal graft in the MSCs group was (11.8±1.6) days,it was significantly longer than (9.6±1.4) days in the PBS control group (P=0.004).The levels of IL-4 mRNA and IL-10 mRNA in the MSCs group were significantly higher than those in the PBS control group (both at P =0.00);while the levels of IFN-γ mRNA and IL-2 mRNA were not significantly different between the groups (both at P＞0.05).The IL-10 protein contents were (22.74 ±7.06),(68.40±12.83) and (215.41 ±44.66)pg/ml in the normal control group,PBS control group and MSCs group,showing significant difference among the three groups (F =55.06,P =0.00) and a significant increase in the MSCs group compared with the PBS control group and the normal control group (both at P ＜ 0.05).Conclusions Subconjunctival injection of MSCs prolongs the survival time of cornea allograft in penetrating keratoplasty probably by modulating the balance between Th1 and Th2 cytokines,especially by up-regulating Th2 cytokines.

Background Corneal transplantation is a primary method for the treatment of serious corneal diseases, but its application is limited because of the shortage of corneal donor.The study on tissue engineering corneal epithelium provides a new approach to corneal transplantation, and the biological scaffold materials for tissue engineering corneal epithelium is an issue of increasing concern.Bacterial cellulose membrane has been used in medical field,but its application in tissue engineering corneal epithelium deserves more researching.Objective This study was to evaluate the biocompatibility of bacterial cellulose membrane as a biological scaffold of tissue engineering corneal epithelium.Methods Corneal epithelium was isolated from 1 month-old New Zealand White rabbit.Corneal epithelial cells were cultured using explant method and identified by detecting the CK-3 expression using immunofluorescence technique.The second generation ceils were inoculated on bacterial cellulose membrane and culture plate, respectively, and the growth status of the cells were examined and compared under the optical microscope.The cell activity/toxicity test was performed by LIVE/DEAD cell staining kit at the third day after inoculation to evaluate the survival rate.The ultrastructure of the cell surface was examined under the scanning electron microscope.The study was performed in accordance with the ARVO Statement.Results Rabbit corneal epithelial cells grew well 1 week after primarily cultured with a cobblestone-like appearance and positive response for CK3 antibody.The cells on the bacterial cellulose membrane presented a round shape and regular arrangement and showed the green fluorescence for LIVE/DEAD test,with the survival rate 100％.Abundant leafy protrusion, microvilli and intercellular junction were seen under the scanning electron microscope.In addition, mitosis phase of cells and many filopodia between the cells and bacterial cellulose membrane were also exhibited.Conclusions Rabbit corneal epithelial cells can grow well in bacterial cellulose membrane.Bacterial cellulose membrane has good biocompatibility, indicating that bacterial cellulose membrane can be used as new biological material for tissue engineering corneal epithelium.

Objective To investigate the effect of cognition training on the recovery of balance after stroke. Methods forty-five hemiplegic strokd patents with cognition deficit were revruited and sddigned into an experimental group and a control group according to the results of an assessment with Loewenstein's Occupational Therapy Cognition Assessment battery (LOTCA). The patients in the two groups folloled routine systmatic rehabilitation programs. The patients in the (LOTCA). The patients in the two groups followed routine systmatic rehabilitation programs.The patiena in the experimental group received pertinent cognition training 5 days/week for 6 weeks. LOTCA,Brunel's Balance Assessment (BBA), the Berg Balance Scale (BBS) and Brunnatrom functional categories were enployed to evaluate the patients before and after treatment with regard to their cognition, balance control and motor function in their lower limbs. Results After treatment, there were statistically significant differences between preand post-treatment BBA and BBS scores in all groups. Average BBA, BBS and LOTCA scores in the experimental group were significantly higher than in the control group. The improvements in BBS, BBA and LOTCA scores in the experimental grou were significantly greater than in control group. Conclusion Cognition treatment can contribute significantly to improving balance and postural control among stroke survivors.

Objective To evaluate the damaged level of auditory processing and diagnose the type of language impairment in an auditory agnosia patient using Psycholinguistic Assessment in Chinese Aphasia (PACA), and predict the outcome of the language function. Methods A patient with a bilateral damage involving the temporal lobe could physically hear the sounds, but was unable to recognize or differentiate between the sounds. Speech and language evaluations were taken with PACA 4 weeks, 8 weeks and 3 months after the onset of disease. Results At 4 weeks, the correct rates were 2.5% and 95.0% in spoken word-picture matching and written word-picture matching (P<0.001), and were 5.6% and 80.6% in spoken semantic knowledge and written semantic knowledge (P<0.001). The scores of auditory input processing function score were low, including phoneme discrimination, spoken word matching with minimal differences, environmental sounds identification. The correct rates were 67.5% in oral picture naming, 0 in word repetition, 70.0% in word reading, 0 in dictation, 100.0% in directly copy writing. At 8 weeks and 3 months, there was complete recovery in written word- picture matching (both 100%) and written semantic knowledge (97.2%, 100%), and improvement in oral picture naming (87.5%, 90%), word repetition (87.5%, 97.5%), picture name writing (77.5%, 87.5%) and copy writing (both 100%) (P<0.05). But no change was found in auditory input processing function examinations, spoken word-picture matching (5.0%, 7.5%), spoken semantic knowledge (2.8%, 5.6%), word repetition (0, 3.3%) and dictation (both 0). Conclusion The patient was diagnosed as auditory agnosia with the inability to distinguish sounds. PACA can well identify the impaired level of auditory comprehension disorder and the outcome of language function recovery for aphasics.

Sixty-nine cases of acute hyperthyroid myopathy admitted from 1990 to 2011 were retrospectively analyzed.Hoarseness was the most common symptom; 17.39％ of the patients had difficulty in breathing; respiratory muscle paralysis and aspiration pneumonia were the main causes of death.Efficient treatment lasting for two weeks was carried out in 81.16％ of the cases.Patients with severe bulbar palsy were often complicated with aspiration pneumonia and thus had high mortality rate.The main treatments in cluded anti-hyperthyroid drugs,corticosteroids,energy supports,and symptomatic management,which usually resulted in good clinical improvement.

Background Reasearches showed that α-melanocyte-stimulating hormone (α-MSH) inhibits inflammation and ameliorates the ocular surface abnormalities in a scopolamine-induced dry eye rat model,and the managing effect of sodium carboxymethylcellulose (CMC) on dry eyes also has been determined.However,whether α-MSH can enhance the therapeutic effects of CMC remains to be investigated.Objective This study was to investigate the protective effects of α-MSH combined with CMC on ocular surface in a scopolamine-induced dry eye rat model.Methods Sixty clean female Wistar rats were randomly divided into normal control group,model control group,NaCl group,CMC group,α-MSH group and α-MSH+CMC group,and 10 rats for each group.The dry eye models were established by subcutaneous injection of scopolamine hydrobromide at 9:00,12:00,15:00 and 18:00 per day for 28 days.0.9％ NaCl solution,1×10 3 mg/ml α-MSH solution,0.5％ CMC eye drop,and 1 ×10-3 mg/ml α-MSH+0.5％ CMC solution were topically administered twice a day (8:00,17:00) since the initial day of modeling according to grouping.Shirmer Ⅰ test (S Ⅰ t),breakup time of tear film (BUT) and corneal fluorescence staining were performed before and 7,14,21,28 days after the application of drugs.At 28 days following the administration of drugs,the eyeballs of the rats were collected.Hemotoxylin and eosin staining was employed to examine the morphology of corneas,and periodic acid schiff (PAS) staining was used to count the conjunctival goblet cells.This study protocol was approved by Experimental Animal Ethic Committee of Tianjin Medical University (SYXK 2009-0001),and the use and care of the rats complied with ARVO Statement.Results The S Ⅰ t and BUT values were significantly reduced,and the corneal fluorescence staining scores were significantly increased over time following modeling in the model control group (all at P＜0.01).No significant differences were found in the S Ⅰ t,BUT and corneal fluorescence staining scores between model control group and NaCl group at various time points (all at P＞0.05).At 7,14 and 21 days after intervention,the S Ⅰ t values were (4.800±0.789),(4.100±0.516) and (4.300±0.856) mm in the α-MSH+CMC group,which were considerably higher than (2.875 ±0.719),(2.375 ±0.619) and (2.532±0.957)mm in the NaCl group (all at P＜0.01).At 7 days after intervention,the BUT values were (4.938± 1.843) seconds and (5.000±1.491) seconds in the α-MSH group and α-MSH+CMC group,which were significantly higher than (3.250±1.000) seconds in the NaCl group (both at P＜0.01).The corneal fluorescence staining scores in the CMC group,α-MSH group and α-MSH+CMC group were significantly lower than that in the NaCl group,with the lowest score in the α-MSH +CMC group (all at P＜0.05).The thickening of corneal epithelial layer,corneal edema and arrangement disorder of corneal stroma were found in the model control group and NaCl group;while slight corneal edema and epithelial cell proliferation were exhibited in the α-MSH+CMC group by hemotoxylin and eosin staining.PAS staining showed that the number of goblet cells was much more in the CMC group,α-MSH group and α-MSH+ CMC group than that in the model control group and NaCl group (all at P ＜ 0.01).Conclusions The sole application of α-MSH or CMC alleviates ocular surface damage and morphological abnormality to certain extent,and the combination of α-MSH and CMC generates more effective protection in comparison with sole administration of α-MSH or CMC.The early application of the drugs plays an improvement role in tear secretion and tear film stability in dry eyes.

Objective To evaluate the colonoscopy surveillance in colorectal cancer patients after surgical removal of the tumor.Methods From June 1986 to June 2007,2762 asymptomatic patients who had underwent operation for colorectal cancer were put into colonoscopy surveillance.They had the first examination 3-6 months after the operation,and were re-examined once a year thereafter for 3 years.The follow-up findings were compared with those from the 218 symptomatic patients who had colorectal cancer surgery from September 1981 to May 1986.Results In 2762 asymptomatic patients,48 cases of multiple primary cancer were detected,in which 39 cases(1.4%) were found at one examination and 9 cases(0.3%)at different examination.The TNM staging of these lesions included stage Ⅰ in 6,stage Ⅱ in 31 and stage Ⅲ in 11.During the surveillance,583 cases(21.1%) of adenoma were detected and endoscopically resected,in which 17(3.2%) were invasive early cancer and 58(9.9%) were high grade dysplasia.In 218 patients with symptoms,29 cases(13.3%) of adenoma and 27 cases( 12.4%) of cancer were detected,including 4 cases of stage Ⅰ cancer,6 of stage Ⅱ and 16 of stage Ⅲ.Conclusion Colonoscopy surveillance in colorectal cancer patients after surgery is important in finding precancerous lesion and early stage cancer,and is recommended in all patients.

Objective To investigate the antimicrobial resistance in the clinical strains isolated from Tongji Hospital to the antimicrobial agents commonly used in 2012.Methods Antimicrobial susceptibility was tested by Kirby-Bauer method.The minimum inhibitory concentration (MIC) of penicillin and ceftriaxone for Streptococcus pneumoniae and vancomycin for Staphylococcus spp.were determined by E-test.All data were analyzed by WHONET 5.6 software.Results A total of 8 191 strains were isolated in 2012,including gram-positive bacteria (2 815,34.4%)and gram-negative bacteria (5 376,65.6%). The top five pathogenic bacteria isolated from outpatients were Escheria coli,Pseudomonas aeruginosa,coagulase-negative Staphylococcus (CNS),Klebsiella spp.and Staphylococcus aureus.For the isolates from non-ICU inpatients,the top five were E.coli,S.aureus,Acinetobacter spp.,Klebsiella spp.and P.aeruginosa.For those isolated from ICU patients,the top five were Acinetobacter spp.,S.aureus,P.aeruginosa,Enterococcus spp.and E.coli.The prevalence of MRSA and MRCNS was 58.1% and 64.3%,respectively.Seventeen strains of vancomycin resistant Enterococcus were identified, including 13 strains of E.faecium with VanA and 4 strains of E.gallinarum with VanA and VanC.The percentage of antimicrobial resistance in E.faecium was significantly higher than that in E.faecalis (P<0.05).A total of 94 strains of carbapenem-resistant Enterobacteriaceae were detected.The prevalence of penicillin-non-susceptible S.pneumoniae in children was much higher than that in adults. The prevalence of carbapenem-resistant P.aerugonosa and Acinetobacter spp. was 28.1% and 56.2% respectively.Beta-lactamase was produced in 41.8% of the H.influenzae and 98.6% of the M. catarrhalis isolates. Conclusions The prevalence of multidrug resistant strains has been increasing, especially vancomycin-resistant Enterococcus and carbapenem-resistant Enterobacteriaceae.

Objective To investigate distribution and drug resistance of pathogenic bacteria in lower respitatory tract infection. Methods Distribution and drug resistance of pathogenic bacteria in lower respitatory tract infection of patients in ICU and non-ICU of our hospital during 2013 were retrospectivly analyzed. The pathogens were identified by manual methods routinely and those difficult to be identified were analyzed by using the VITEK-2-COMPACT instrument. Antimicrobial susceptibility of these isolates were tested by Kirby-Bauey methods routinely. Results In total, 956 strains were isolated from lower respitatory tract infection of patients in ICU, including 231 strains of gram-positive cocci (24. 2%), 680 strains of gram-negative bacteria (71. 1%), 45 strains of fungi (4. 7%). In patients of non-ICU, 4 464 strains were isolated, including 1 090 strains of gram-positive cocci (24. 4%), 3 226 strains of gram-negative bacteria (72. 3%), and 148 strains of fungi (3. 3%). Staphylococcus aureus, acinetobacter baumannii and pseudomonas aeruginosa were the most frequent isolates in patients of ICU and non-ICU. The overall prevalence of methicillin resistant staphylococcus aureus (MRSA) in staphylococcus aureus was 87. 0%in ICU and 74. 0% in non-ICU. MSSA was sensitive to the most antibiotics ( more than 80. 0% of the strains were sensitive to common antibiotics) except penicillin, erythromycin and clindamycin. MRSA was sensitive to trimethoprim-sulfamethoxazole and fosfomycin (more than 75. 0% of the strains were sensitive to the antibiotics) except for vancomycin, teicoplanin and linezolid. Acinetobacter baumannii was more resistant to the antibiotics (less than 40. 0% of the strains were susceptible to the antibiotics). Pseudomonas aeruginosa from ICU was more resistant to the antibiotics ( less than 50. 0% of the strains were sensitive to the antibiotics) than that from non-ICU. Stenotrophomonas maltophilia was sensitive to trimethoprim-sulfamethoxazole, levofloxacin and minocycline (more than 80. 0% of the strains were sensitive to the antibiotics). Escherichia coli and Klebsiella pneumoniae were sensitive to Piperacillin-tazobactam and Amikacin except for meropenem and imipenem ( more than 80. 0% of the strains were sensitive to the antibiotics) . Conclusion Gram-negative bacteria was the most frequent isolate in lower respitatory tract infection of our hospital during 2013. Staphylococcus aureus, acinetobacter baumannii and pseudomonas aeruginosa were the most frequent isolates in ICU and non-ICU. Resistance to the antibiotics was more common in ICU than in non-ICU. Antibiotics should be prescribed according to bacterial resistance results reasonably in order to prevent the spread of drug-resistant strains.

Objective To analyze the correlation between the dry eye symptom and signs in middle-aged and aged people by Lipiview(R) ocular surface interferometry.Methods A total of 170 eyes from 85 middle-aged and aged people (55-80 years old) were examined with Standard Patient Evaluaion of Eye Dryness(SPEED) questionnaire and Lipiview(R) ocular surface interferometry,meanwhile obtaining the consent of the subjects.Then the correlation between the symptom and Lipid Layer Thinckness (LLT) and partial blink rate were analyzed.Results SPEED questionnaire score was 6.13 ±4.60.LLT was(74.88 ± 21.16) nm.LLT variability was 4.47 ± 3.40.The partial blink rate was 0.62 ± 0.36.The Spearman correlation analysis indicated that the nega-tive correlation was shown between the SPEED questionnaire score and LLT (r =-0.823,P =0.000),SPEED questionnaire score and LLT variability(r =-0.268,P =0.018).The positive correlation were found between LLT and Std.Dev of LLT (r =0.339,P =0.030).However,the results of the SPEED score were uncorrelated with partial blink rate (P ＞ 0.05).Conclusion The results from this study demonstrate that correlation among symptom,LLT and LLT variability,but poor correlation between symptom and partial blink rate.