Background Researching the pathological characteristics and components of cells in internal limiting membrane of idiopathic macular hole (IMH) has an important clinical significance for the prevention of IMH.However,the study results are still disputable.Objective This study was to investigate the histopathological features of internal limiting membrane of IMH and the types of cells inside it, and explore the pathomechanism of IMH.Methods Seven specimens of internal limiting membrane were obtained during the vitrectomy with IMH patients in the Second Affiliated Hospital of Guangzhou Medical University from February 2012 to August 2013 under the informed consent of patients.The histopathological examination was performed for the structural observation and cellular identification of internal limiting membrane.The expression and location of glial fibrillary acidic protein (GFAP), CD45 and CD44 in internal limiting membrane were examined by using immunochemistry and immunofluorescence technology.Results All the seven specimens showed continuous undulating membrane with red staining.Two specimens appeared to be uniform in thickness and few cells were distributed in the specimens.The internal limiting membranes were uneven in thickness in the other specimens with retinal pigment epithelial cells,neuroglia cells,fibrocytes,macrophages and lymphocytes in them.Immunochemistry showed the positive expression of GFAP in the outer layer of the specimens.CD45 positive cells were detected in the internal limiting membranes, and CD44 was detected in the inner layer of the specimens.Conclusions Few cells exist in the internal limiting memranes of IMH.However,neuroglia cells and CD45 positive cells emerge in the internal limiting memranes of stage 3 or above IMH eyes, indicating the proliferation of cells and immuno-inflammation response exist during the IMH development.The up-regulation of CD44 expression promotes inflammatory response of internal limiting memranes.

Objective:To study the instant and short term effects of diagnostic ultrasound on ultrastructure and hydrogen peroxide cytochemistry of human villi. Methods: Fifteen healthy women with gestational ages of 6 to 8 weeks were divided into 4 groups. Group A( n =3),B( n =4),C( n =4) and D( n =4) were exposured to diagnostic ultrasound for 0,10,20 and 20 min respectively. In group A, B, and C, the villi were taken out immediately after ultrasound exposure and were studied. In group D, the villi were taken out 3 d after ultrasound exposure. Results: The results showed that there were changes only in group C. Enlargement of endoplasmic reticulum and mitochondrial intracristal space were observed in syntrophoblast cells. In group A, B, and D, there were no evident abnormality. Conclusion: The conventional acoustic exposure of diagnostic ultrasound is safe for human villi.

ObjectiveTo investigate the diagnosis and the treatment of male interstitial cystitis (IC) to improve the efficiency.MethodsEighteen cases of IC male patients treated from Jan 2010 to Dec 2010 who suffered from suprapnbic pain urinary frequency and urgency were analyzed retrospectively.All these patients were misdiagnosed as category Ⅲ chronic prostatitis.According to the NIDDK diagnostic criteria of IC,Pelvic Pain and Urgency Frequency (PUF) scoring,potassium sensitivity test (PST),and cystoscopy under anaesthesia were used to establish the diagnosis of IC.24 h urinary diary,routine uronoscopy,prostate fluid routine and bacterial culture examination were taken before the treatment of hydrodistention and intravesical instillation of heparin.ResultsAfter the follow-up 12 to 25 months ( average,19 months),the symptoms improved distinctly.The PUF scoring was 19.2 ±4.1 before treatment and 13.6 ±2.4 after treatment respectively ( P ＜ 0.01 ).24 hours' frequency and amount of urination were (7.5 ± 4.3)times and (241.7 ±45.3) ml after treatment compared with (11.5 ±3.9) times and (159.5 ±30.8) ml before treatment ( P ＜ 0.01 ).ConclusionsThe male IC and chronic prostatitis share the same symptoms.They can be differentiated by the IC diagnosis.The treatment of hydrodistention alone with oral tolterodine tartrate sustained release tablets and intravesical instillation of heparin can evidently improve the symptoms of the male IC patients.

Background Recent studies indicated that human amnion mesenchymal stem cells (hAMSCs) can be induced to differentiate into multiple types of cells in vitro,but whether the hAMSCs can differentiate into ocular surface cells has not been reported yet.Objective This study was to investigate the feasibility of inducing differentiation of hAMSCs into ocular surface cells by co-culturing with human bulbar conjunctiva fibroblasts (hBCFs).Methods This study was approved by Ethic Committee of Affiliated Second Hospital of Guangzhou Medical University.HAMSCs were isolated from placenta under the informed consent of healthy delivery women.hAMSCs were cultured,passaged and identified by detecting the expressions of CD44,CD45,CD73,CD90 in the cells with flow cytometer,osteogenesis and adipogenic differentiation experiments.Human conjunctival tissue was obtained during the eye operation under the informed consent of patients and hBCFs were isolated and cultured with explant culture.The cells were divided into the hAMSCs culture group and the hAMSCs and hBCFs co-culture group and cultivated in Transwell chambers for 7 days.The expressions of cytokeratin 19 (CK19) and α-smooth muscle actin (α-SMA) in the cells were assayed by immnofluorescence technique.Results Cultured hAMSCs showed the slender shape and cell body enlarged with passage.CD44,CD73 and CD90 were expressed in the cells,and the expression of CD45 was absent.After 3-4 weeks of osteogenesis and adipogenic induce,the cells showed red staining for alizarin and oil red O.In the co-culture group of hAMSCs and hBCFs,hAMSCs presented the epithelioid cell-like in shape and showed the positive response for CK19 and weaker response for α-SMA.However,in the hAMSCs culture group,the cells showed the positive response for α-SMA and absent response for CK19.Conclusions The hAMSCs can differentiate into ocular surface cells after being induced by hBCFs.And the differentiation mechanism is possibly relevant to mesenchymal cells epithelium.

Background It has not been reported that if the visual cortex M receptor changed during the development of myopia and how it changed if given acupuncture treatment.Objective The aim of this study was to observe the effect of electroacupuncture stimulation on the expression of acetylcholine receptors M1 (AchRM1) in visual cortex of guinea with lens-induced myopia (LIM).Methods Forty-eight three-week-old healthy guinea pigs were randomized into the normal control group,the LIM model group and the LIM electroacupuncture group.The right eyes of the guinea pigs were selected as the experimental eyes.LIM was created by monocularly wearing of-10 D lens for 4 weeks in the right eyes in the LIM model group and LIM electroacupuncture group,and then the acupuncture at the temple and hegu point was performed for 30 minutes per day for consequent 4 weeks,in the LIM electroacupuncture group.The fellow eyes of the guinea pigs were used as the self-control eyes.The refractive power and axial length were examined with retinoscopy and A-type sonography before and 4 weeks after modeling,respectively.The animals were sacrificed by excessive anesthesia at the fourth week after acupuncture and visual vertex tissue was obtained.The expression of M1 receptor mRNA in visual vertex was detected by fluorescence quantitative PCR,and the content of M1 receptor protein in visual vertex was assyed by ELISA.The study protocal was approved by Animal Ethic Committee of Shandong University of Traditional Chinese Medicine,and the use and care complied with Statement of the Association for Research in Vision and Ophthalmology.Results At the fourth week after modeling,the mean diopters were (-3.24±0.28) D and (-3.30±0.45) D in the LIM model group and the LIM eleetroacupuncture group,which were significantly higher than (0.83 ±0.86)D in the normal control group (both at P=0.000),and there was no significant difference in the diopter between the LIM model group and the LIM electroacupuncture group (t =0.200,P =0.659).The mean axial lengths were (8.67 ±0.14) mm and (8.60±0.06) mm in the LIM model group and the LIM electroacupuncture group,which were considerably increased in comparison with (8.33±0.08)mm in the normal control group (both at P＜0.05).The relative expression levels of AchRM1 mRNA in visual cortex were 0.79±0.18,1.36±0.23 and 1.13±0.13 in the normal control group,LIM model group and LIM electroacupuncture group,and the relative expression level of AchRM1 mRNA in the LIM electroacupuncture group was significantly higher than that of the normal control group and lower than that of the LIM model group (both at P＜0.05).In addition,the contents of AchRM1 receptor protein in the visual cortex were 248.00±33.31,455.17±42.40 and 396.17±47.57 in the normal control group,LIM model group and the LIM electroacupuncture group,with a similar pattern among the groups (both at P＜0.05).Conclusions A electroacupuncture stimmulation do not affect the myopic diopter and axial length in LIM model.The AchRM1 and AchRM1 receptor in the visual cortex up-regulate in LIM eyes,infering that electroacupuncture stimmulation can improve vision by decreasing the level of AchRM1 receptor in visual cortex in LIM eyes in guinea pigs.

BACKGROUND:Conditioned medium from mesenchymal stem cels (MSC-CM) may represent a promising alternative to MSCs transplantation. Previous studies have shown that inflammatory activation can strengthen the multiple biological potencies of MSCs; however, normal MSCs with insufficiency of immunocompetence and migration ability are not effective for tissue damage repair. OBJECTIVE:To investigate differential effects of MSC-CM with and without inflammatory activation on radiation-induced intestinal injury.METHODS:MSCs from the bone marrow of SD rats were separated, cultured and identified, and then co-cultured with non-irradiated IEC-6 or irradiated IEC-6 in a transwel system for 24 hours. Then, MSCs with inflammatory activation were cultured alone for another 48 hours. After that, the supernatant was colected as non-activated MSC-CM (MSC-CMNOR) and MSC-CM under radiation-induced inflammatory condition (MSC-CMIR). Rats were exposed to 14 Gy whole abdominal irradiation and randomly divided into four groups: control group, radiation injury group (DMEM/F12), MSC-CMNOR group and MSC-CMIR group. Continuous administration was givenvia tail vein and intraperitoneal implantation of Alzet microosmotic pumps. Intestinal samples were colected at 1, 3, 7 days after radiation for analysis of short circuit variation, at 3 days after radiation for analysis of intestinal epithelium ultrastructure, and at 1, 3, 5, 7, 14 days after radiation for histological observation of the intestinal epithelium using hematoxylin-eosin staining. Blood samples were colected at 1, 3, 7 days after radiation for analysis of serum xylose levels. In addition, the survival state and survival time of rats were observed and recorded. RESULTS AND CONCLUSION: The short circuit variation responding to electrical field stimulation was significantly reduced at al frequencies, but it was significantly improved in the MSC-CMIR group. Similarly, the intestinal absorption (serum xylose levels) was also significantly impaired by irradiation, but improved by delivery of MSC-CMIR (P < 0.05). At 3 days after MSC-CMIR infusion, the intestinal epithelium exhibited an increase in crypt size and vilous length (P < 0.05). Under the electron microscope, a reduction in intestinal microvili and open tight junctions in irradiated intestinal epithelium was found, and the intestine from rats treated with MSC-CMIR had more obvious tight junctions. In addition, treatment with MSC-CMIR dramaticaly improved the survival rate and mean survival time of irradiated rats as compared to those treated with DMEM/F12 or MSC-CMNOR (P < 0.05). Taken together, the present study demonstrated that MSC-CMIR , but not non-activated MSC-CM, improves the structural and functional restoration of the smal intestine after radiation-induced intestinal injury.

AIM:To observe the expression of nuclear factor-kappa B ( NF-κB) , N-methyl-D-aspartic acid re-ceptor 2B (NR2B) and inducible nitric oxide synthase (iNOS) in the spinal cord in a rat model of chronic constriction in-jury (CCI) of the sciatic nerve.METHODS:Fifty-six adult male Sprague-Dawley rats weighing 180~220 g were random-ly divided into sham group (n=8) and CCI group (n=48).The mechanical withdrawal threshold (MWT) and paw with-drawal latency (PWL) of the hind paws were measured 1 d before CCI and 1 d, 4 d, 7 d, 14 d and 21 d after surgery.The L4~L6 segment of the spinal cord was taken for determining the expression of NF-κB, NR2B and iNOS by RT-PCR and Western blotting.RESULTS:At 1 d, 4 d, 7 d, 14 d and 21 d after surgery, the MWT and PWL in CCI group were obviously lower than those in sham group .The expression of NF-κB, NR2B and iNOS at mRNA and protein levels in-creased significantly.Positive correlations were found between the mRNA expression of NF-κB and iNOS (r=0.842, P<0.05), and between the mRNA expression of NR2B and iNOS (r=0.833, P<0.05).CONCLUSION:The generation and maintenance of hyperalgesia in sciatic nerve injury rats may attribute to the activation of NF -κB and NR2B and concom-itant increase in iNOS .

Objective To explore the changes of proteomic spectra from plasma of patients with lung cancer or benign lung diseases and health controls in order to establish a primary diagnosis model of lung cancer. Methods The proteomic spectra from plasma of 108 patients with lung cancer, 40 patients with benign lung diseases and 22 healthy individuals were analysed by surface-enhanced laser desorption/ionization time of flight mass spectrometry ( SELDI-TOF-MS). The best decision tree model was established by cluster analysis and principal component analysis. Then the model was blindly validated by the protein of 21 patients with lung benign diseases and 47 patients with stage I lung cancer. Results Twenty-three significantly differentially expressed protein peaks were successfully detected (P <0.001). Blinded validation suggested that the accuracy for diagnosing lung cancer was 72. 06%, the sensitivity and specificity were 72. 34% and 71.43%, respectively, and the positive predictive value and negative predictive value were 85. 0% and 78. 95%, respectively. Conclusion SELDI-TOF-MS protein chip technology provides a new tool for the early diagnosis of lung cancer.

OBJECTIVEThe effect of chronic morphine treatment on the I(A) (transient outward K+ current) of prefrontal cortical neurons of newborn rat. On this basis, we use AGS3 antibody to inhibit the function of AGS3, for observing the impact of AGS3 on the I(A), thus further explore the mechanism of AGS3 protein in morphine addiction.

METHODSBy using whole-cell patch-clamp technique, I(A) was recorded. In the whole-cell configuration, observed the impact of morphine on the current density-voltage curve (I-V) of I(A) and the effect of AGS3 antibody with three different concentrations on the I(A) of morphine treated rat prefrontal cortical neurons.

RESULTSMorphine increased the I(A). When the test potential was + 55 mV, different concentrations of AGS3, 10(-3) microg/L, 10(-2) micdrog/L and 10(-1) microg/L acted on morphine treated rat prefrontal cortical neurons, the enhanced IA by morphine was inhibited.

CONCLUSIONMorphine increases the I(A), AGS3 protein may participate in signal transduction pathway involved with I(A).

Animals ; Animals, Newborn ; Antibodies, Monoclonal ; pharmacology ; Carrier Proteins ; immunology ; metabolism ; Female ; Male ; Morphine ; adverse effects ; Neurons ; metabolism ; Patch-Clamp Techniques ; Potassium Channels ; drug effects ; Prefrontal Cortex ; metabolism ; Rats ; Rats, Sprague-Dawley ; Substance-Related Disorders ; metabolism ; physiopathology

Actins ; metabolism ; Animals ; Male ; Morphine ; adverse effects ; Morphine Dependence ; metabolism ; Prefrontal Cortex ; metabolism ; Proteome ; metabolism ; Rats ; Rats, Sprague-Dawley ; Substance Withdrawal Syndrome ; metabolism ; Synaptosomal-Associated Protein 25 ; metabolism