AlM:To analyze and compare the effect of femtosecond laser micro - incision corneal stromal lens excision ( SMlLE) and excimer laser in situ keratomileusis ( LASlK) in the treatment of myopia after operation, to explore the safety, operability and prediction of SMlLE.METHODS:ln this prospective clinical controlled study, 100 cases ( 200 eyes ) received SMlLE and 100 cases ( 200 eyes) undergone LASl in our hospital in the same period were selected. Uncorrected visual acuity, diopter, corrected visual acuity, slit lamp examination, intraocular pressure and corneal anterior segment OCT, corneal topography (Obscan ll) of two groups in 1d, 1wk, 1, 3, 6mo, 1a were compared. lndependent samples t test was used for data analysis.RESULTS:1) Postoperative slit lamp examination:after 1d in SMlLE group, there were less eyes had corneal layer between mild cloudy or edema; postoperative 1wk corneal layer disappeared, cornea became clear and transparent. 2 ) Postoperative vision recovery: 1d after operation, vision recovery in LASlK group was better than that in SMlLE group, the difference was statistically significant (P<0. 01), there were no significant differences at 1wk, 1, 3, 6mo, 1a after operation ( P>0. 05 ). 3 ) Obscan ll examination: graphics in the SMlLE group was more regular and placed in the center, no eccentric and irregular graphics, better than that in the LASlK group. 4) Anterior segment OCT examination:postoperative corneal flap in the SMlLE group was more uniform and accurate, but it was thin in the center and slightly thick the peripheral part in the LASlK groups. 5 ) Postoperative visual quality assessment used subjective questionnaire survey. The two groups had statistically significant difference on 4 points and 1 points (P<0. 05). Complains in the LASlK groups were more that that in the SMlLE group. While, no complain of the SMlLE group was higher than that of the LASlK group. Glare of postoperative patients with night vision and dark environment in the SMlLE group was better than that of the LASlK group.CONCLUSlON: SMlLE is safe, effective, stable and predictable for the correction of myopia.

Calcinosis ; pathology ; surgery ; Child ; Diagnosis, Differential ; Humans ; Knee ; Male ; Skin Diseases ; pathology ; surgery

Objective To explore the value of prenatal genetical diagnosis by mutation analysis of achondroplasia (ACH) fibroblast growth factor receptor 3 (FGFR3) gene.Methods Genomic DNA from nine ACH patients and their parents in Gansu Maternal and Child Health Hospital from July,2010 to December,2014 was prepared for polymerase chain reaction.Direct sequencing revealed the samples were performed after amplification of exon 10 of FGFR3 containing the potential mutation.Fetal DNA was extracted from cells in both amniotic fluid and umbilical cord,and then exon 10 of FGFR3 was also tested.Three fetuses with short-limb dysplasia were also included and prenatal diagnosis was offered to them through amniocentesis or cordocentesis.Results Prenatal ultrasonography test showed shorter femoral length,which was less than 2-3 standard deviation of normal reference dysplasia fetal performance for femoral short.Femur length is lower than 2-3 standard deviation minus normal value,and discrepancy in biparietal diameter compared with fetuses at the same gestational age.In the four families with one ACH parent,c.1138G ＞ A heterozygous mutation was detected in all of the four mothers,while two fetuses among them showed c.1138G ＞ A heterozygous mutation mutation and the other two were normal.There were other two fetuses with c.1138G ＞ A heterozygous mutation from other two families,one's father had c.1138G ＞ A heterozygous mutations,but not the mother,the other had c.1138G ＞ A heterozygous mutations in both the mother and father.Among the three families with unaffected parents but each had a de novo c.1138G ＞ A mutation child,no mutation of c.1138G ＞ A genotype was detected in their fetuses,neither in the three fetus with short limb dysplasia.Four fetuses with a c.1138G ＞ A mutation and three with short-limb dysplasia were terminated.The other five fetuses whose genotype was normal were born and healthy with normal phenotype at one-year-old follow-up.Conclusion FGFR3 genetic analysis could provide information for genetic counseling and prenatal diagnosis for ACH parents or parents who had an ACH baby to prevent birth defect.

Objective To investigate the change of matrix metalloproteinase-9 (MMP-9) and tissue inhibitor of metalloproteinase-1 (TIMP-1) in the skin of diabetic rats, and to explore the potential role of MMP-9/TIMP-1. Methods Diabetic rats were induced with streptozotocin (STZ). Then all rats were maintained for 6 weeks. Routine pathological examination and immnnohistochemistry were made to reveal the histological and cytological appearances. RT-PCR and Western blotting were used to detect the expression of mRNA and protein of MMP-9 and TIMP-1 in the skin. Results Six weeks after STZ treatment, examination of HE-stained skin sections from normal and diabetic animals revealed that the epidermis and dermis layers were thinner in diabetic rats than those in control rats. The skin of diabetic rats showed features of atrophy such as disorganization of connective tissue fiber bundles and enlarged space between collagen fiber bundles. In contrast, thick bundles of connective tissue were observed in the dermis of normal rat skin. In normal skin, cells had a bipolar, spindle-shaped appearance in the thick collagen bundles, while in the skin of diabetic animals the interstitial cells had a rounded, shrunken and crenated appearance. The relative values of expression of MMP-9 in diabetic group were higher than those in normal group with significant difference, however, the relative values of expression of TIMP-I in diabetic group were lower than those in control group. Conclusion The changes in cutaneous histology and cytology appear earlier than skin wound. These "underlying disorders" may be associated with the imbalance of MMP-9/TIMP-1.

To study the contribution of T cell subsets in the pathogenesis of Murine hepatitis virus Type3 (MHV-3) induced chronic viral hepatitis in C3H/Hej mice, ninety C3H/Hej mice were chosen to individually receive 10 plaque forming units (PFU) of MHV-3 intraperitoneally. The changes of virus titer and pathology in liver tissue were examined by standard plaque assay and by the hematoxylin/eosin (HE) staining method from 2 days post MHV-3 infection. The ratios of T cell subsets including CD3+CD4+CD8-, CD3+CD4-CD8+, CD3+CD4-CD8-, CD3+CD4+CD25+, CD3+CD4+CD25- and CD3+CD4-CD25+ T lymphocyte of total T lymphocytes in blood, spleen and liver were examined at 0, 2, 4, 6,8, 10, 12, 15, 20, 25, 30, 40 days post MHV-3 infection by flow cytosorting. We observed that the virus titer raised and showed persistent virus duplications and inflammatory changes in the livers of C3H/Hej mice from 2 days post MHV-3 infection. The double negative T cell (DN Treg cell) and CD4+CD25+ T cell ratios increased significantly from 2 days post MHV-3 infection in C3H/Hej mice, and CD3+CD4+CD8-, CD3+CD4-CD8+, CD3+CD4+CD25- and CD3+CD4-CD25+ T cell ratios decreased accordingly. In conclusion, the changes of virus titer and pathology in the livers of C3H/Hej mice post MHV-3 suggest their contribution to viral persistence. Further characterizations of DN Treg cells are that infection indicates that MHV-3 could induce the chronic inflammation in livers of C3H/Hej mice.The increase of the DN Treg cell and CD4+CD25+ T cell ratios in C3H/Hej mice post MHV-3 infection suggests that DN Treg cells and CD4+CD25+ T cells may both have important suppressive immunomodulation functions in the development of chronic viral hepatitis and have important roles in the virus persistent infection. Further characterizations of DNT cell and CD4+CD25+ T cell are under investigation.

Background Femtosecond laser in situ keratomileusis (LASIK) inevitably injury keratocytes and corneal nerve fibers.The research report about postoperative morphological changes of corneal nerve regeneration and keratocytes in femtosecond LASIK is still rare.Objective The aim of this study was to observe the kinetic changes of keratocytes and corneal nerve in corneal flap after femtosecond LASIK.Method Femtosecond laser manufacture corneal flap of LASIK surgery was performed on 60 eyes of 30 patients with refractive error using both femtosecond laser system and excimer laser treatment system.The repair of corneal wound was examined by slit lamp microscope,and the morphology of keratocytes and corneal nerve were observed with confocal microscope 1 week,1month,3 months after surgery,respectively.Results No haze or flap folds were found under the slit lamp microscope from 1 week through 3 months after operation.One week after surgery,the corneal stromal cells at the interface of the corneal flap appeared to be a mild activation status in 42 eyes (70％),but the activated cells gradually reduced with lapse of time.Three months after surgery,mild activation state still was found in 7 eyes (12％).One week after surgery,independent,short (＜50 μm),curved subbasal nerve fibers were exhibited in 7 eyes (12％),and curved filamentous nerve fibers were discovered in 48 eyes (80％) one month after surgery.The nerve fiber length of subbasal nerve was ＞200 μm in 27 eyes (45％) and classes beaded structure appeared 3 months after operation but were still different with preoperative subbasal nerve fibers.One week after the operation,filaments or discontinuous nerve fibers could been seen in 46 eyes (77％) at theinterface,and long nerve fibers or filamentous nerves were visible around the terminal or periphery of nerve fibers in 49 eyes (82％) one month after surgery,and long nerve fibers or filaments of nerve fibers were visible in 57 eyes (95％) 3 months after the surgery.Conclusions Femtosecond LASIK cause wound reaction at cellular level.Corneal nerve fibers recover with the extension of time,but there are still some morphological differences 3 months after surgery from preoperation.

ObjectiveTo explore the mechanism of extracorporeal shock wave (ESW) in treating osteogenic disorders and the ideal energy level. MethodsAfter success in marrow aspiration from patients' iliac crest, hMSCs were isolated by Percoll density gradient centrifugation and cultured in Dulbecco's modified Eagle's medium in a 5% CO2 and 37 ℃ incubator. Optimal ESW dose was determined by MTT of kinase-marked cytobiology. After hMSCs were exposed to ESW, their morphocytologic change, rate of adherence and doubling time were observed with IPCM. Enzyme cytochemistry reaction for the activity of alkaline phosphatase was also examined. ResultsESW of 5 kV and 100 times could increase cells' viability and proliferation (P<0.01), but higher than 7 kV would inhibit them. Rate of adherence of hMSCs in exposure group of passage 5 reached to 61.54%, which was significantly different from control group(P<0.05). Compared with control group, the MSCs' doubling time was short for 1.72 d (P<0.05). The curve of normal alkaline phosphatase activity of hMSCs was like type S, but ESW shortened its latent period, and promoted its peak time, which was significantly different from control group.ConclusionESW of 5 kV and 100 times can optimally promote the proliferation and activity of osteogen of hMSCs in vitro.

Objective To evaluate the value of CT guided percutaneous transthoracic needle biopsy (PTNB) for the diagnosis of lung ground‐glass opacity (GGO) with a Meta‐analysis .Methods Relevant English and Chinese language studies were searched on the PubMed ,EMBASE ,EBSCO ,OVID ,CNKI ,CBM ,VIP and WANFANG databases ,respectively .Data were calculated with software of Stata 12 .0 and Meta‐Disc 1 .4 .Results 6 of 82 retrieved studies were included (n=341) .The pooled sensitivity ,specifici‐ty ,LR+ ,LR- ,DOR ,AUC and 95% CI were 0 .92(0 .86-0 .95) ,0 .98(0 .85-1 .00) ,49 .03(5 .72-420 .18) ,0 .08(0 .05-0 .15) , 586 .24(65 .18-5 272 .83) and 0 .99(0 .98-1 .00) ,respectively .Conclusion CT guided PTNB can be used as one of the primary examination modalities for lung GGO with moderate sensitivity and specificity .

This study was aimed to investigate the effect of CD44 gene silence on the drug resistance and biologic activity of human multidrug resistant leukemia cell line K562/A02. The oligonucleotides of CD44 gene were designed according to related data of GenBank, double-stranded DNA was produced by annealing, and was inserted into pGCsilencerU6/Neo/GFP vector. The resultant recombinant plasmid pGCsiRNA-CD44 was transfected into K562/A02 cell line. Expressions of CD44, mdr-1 and blc-2 mRNA were assayed by real time RT-PCR. The 50% inhibitory concentration (IC50) of doxorubicin (ADM) for K562/A02 cell line was determined by MTT method. Cell cycle was determined by flow cytometry. The morphology of apoptotic cells was examined by Hochst 33258 staining. The results indicated that the siRNA eukaryotic plasmid directing at CD44 gene could effectively silence the CD44 gene of K562/A02 cells; as compared with control group, the CD44 expression in K562/A02 cells transfected with 4 pGCsiRNA-CD44 plasmids was obviously inhibited, while the inhibition of CD44 expression in cells transfected with siCD44-1 was strongest. After being transfected with pGCsiRNA-CD44, the expression of CD44 mRNA in K562/A02 cells reduced by 64.1% (p<0.05), at the same time the expression of mdr-1 and bcl-2 mRNA in pGCsiRNA-CD44-transfected K562/A02 cells reduced by 25.6% and 50.8% respectively. IC50 of K562/A02 cells after transfection decreased to (8.77+/-1.63) microg/ml and was obviously lower than that of control (17.97+/-1.61) microg/ml (p<0.01). After transfection for 48 hours, the ratio of K562/A02 cells in G0/G1 increased by 10.7%, and the cells displayed karyopyknosis, nuclear margination and apoptotic bodies. It is concluded that the siRNA plasmid specifically targeting CD44 gene can remarkably down-regulate the expression of CD44 gene, inhibit K562/A02 cell proliferation, induce its apoptosis and effectively reverse the multidrug resistance of K562/A02 cells.
Apoptosis ; Cell Line, Tumor ; Cell Proliferation ; Drug Resistance, Multiple ; genetics ; Drug Resistance, Neoplasm ; genetics ; Gene Silencing ; Genetic Vectors ; Humans ; Hyaluronan Receptors ; genetics ; K562 Cells ; RNA, Small Interfering

OBJECTIVETo investigate the effect of DNA methylation in combination with histone deacetylase inhibitor on transcription regulation of Ras associated domain family gene 1(RASSF1A) tumor suppressor gene and the molecular biological behaviors in U266 cells.

METHODSThe U266 cells were treated with different doses of 5-Aza-2'-deoxycytidine (5-Aza-CdR) and Valproate (VPA) each alone or in combination. Methylation-specific PCR (MSP) was used to detect CpG island methylation in RASSF1A promoter. Quantitative real-time reverse transcription polymerase chain reaction (RQ-PCR) was used to examine the expression of RASSF1A gene in U266 cells. MTT was used for cell proliferation. Cell apoptosis and cell cycle were analyzed by flow cytometry.

RESULTSThe methylation of RASSF1A gene promoter was detected in U266 cells, while there was little RASSF1A gene expressing in the control group. The demethylation effect could be detected in the 5-Aza-CdR treated and combined treatment groups but no in the VPA group. The expression level of RASSF1A was induced by 5-Aza-CdR in a concentration-dependent manner while VPA had no such effect. The expression level of RASSF1A mRNA was increased significantly in the combined treatment group. Higher growth inhibition and apoptosis effects were found in 5-Aza-CdR and VPA combination group than that in 5-Aza-CdR or VPA alone group (P < 0.05). After treatment with 5-Aza-CdR or VPA alone for 72 h, more cells were arrested in G(0)/G(1) phase as conpared with control group (P < 0.05), and even more cells were so arrested in combined treatment group (P < 0.05).

CONCLUSIONDNA methylation and histone deacetylase inhibitor can synergistically induce demethylation of the RASSF1A gene, re-express RASSF1A gene silenced in U266 cells, inhibit the proliferation of U266 cells and induce cell apoptosis.