Background The injury or surgery of cornea cause the proliferation of corneal stromal cells and scar formation.Recent research showed that cureumin can obviously reduce the degree of fibrosis of tissue.But if curcumm play inhibitory effect on corneal keratocytes fibrosis is rarely reported.Objecttve This studv was to investigate the effect of curcumin on the transformation of corneal keratocytes into fibroblasts in vitro and further explore the antifibrotic effect of curcumin on corneal keratocytes.Methods The murine corneal keratocytes from 150 BALB/c mice were isolated and primary culture in DMEM culture medium containing 10% fetal bovine serum and then divided into blank control group(inducer group,CG),low-dose group(CG+7.5 mg/L curcumin),mediumdose group(CG+10.0 mg/L curcumin),high-dose group(CG+12.5 mg/L curcumin),non-inducer group.Seven days following intervention,the expression of cell markers such as keratocan,aldehyde dehydrogenase(ALDH),decorin and fibronectin-1 in keratocytes were analyzed by RT-PCR.The effect of curcumin on cultured murine corneal keratocytes proliferation was evaluated by MTS technique.The expression of fibronectin-1 in murine cornea was investigated by immunofluorescence assay.Results The primarily cultured keratocytes showed tlIe fusiform-like shape with the abundant cytoplasm and big nuclei.In the presence of curcumin,the mRNA levels of keratocan and ALDH were down-regulated and those of CD90 and decorin were up-regulated,showing the significantly differences with the increase of dose(P<0.05),but the expression pf fibronectin-i was not obviously changed with the alteration of dose of curcumin. MTS showed that the inhibitory rates of curcumin on keratocytes in 10.0 mg/L and 2. 5 mg/L groups were enhanced in comparison with 7.5 mg/L group, showing statistically significant difference among three groups( F = 956.00, P<0.05). The expression of fibronectin-1 was found in the corneal keratocytes with the red fluorescence in stroma. Conclusion Curcumin can inhibit the fibrosis of corneal keratoeytes in a dose-dependent manner. These results offer a preliminary theoretical basis for the application of curcumin in controlling corneal scar formation during wound healing.

Background Researches demonstrated that dendritic cells(DCs) are uniformly immature in the central cornea but mature in the peripheral region of cornea.So an important question is which factor impact the maturation of DCs,especially in terms of corneal transplant rejection and the known roles of DCs in the development and persistence of some corneal diseases.Objective This study aimed to examine whether corneal stroma cells (CSCs) inhibit DCs maturation through secreting transforming growth factor beta 2 (TGF-β2) and prostaglandin E2 (PGE2).Methods DCs,T cells and CSCs were isolated and cultured from clean BALB/c and C57BL/6 mice.The level of PGE2 and TGF-β2in CSCs culture supernatant and the fresh RPMI 1640 medium were then analyzed by enzyme linked immunosorbent assay (ELISA).During the DCs maturation stage,the neutralizing TGF-β2 antibody and the EP2 receptor antagonist AH6809 were added in the CSCs culture supernatant respectively.According to the different treatment,cultured cells were assigned to different groups as follows:control group,CSCs culture supernatant group,AH6809 group,TGF-β2 antibody group,AH6809 +TGF-β2 antibody group.Subsequently,the cellular surface markers for DCs,including CD11c,CD80,CD86,and MHC- Ⅱ,were analyzed by flow cytometry.The capability of stimulating the proliferation of T lymphocytes was evaluated by allogeneic mixed lymphocyte reactions,and the function of endocytosis was assessed by fluorescein isothiocyanate-dextran(FITC) uptake.Results The data of ELISA showed a higher concentration of TGF-β2 and PGE2 in murine CSCs culture supernatant than in the fresh RPMI 1640 medium.Compared with the CSCs culture supernatant group,the expression of CD80,CD86,and MHC- Ⅱ was up-regulated ( P ＜ 0.05 ),the expression of dextran was down-regulated ( P ＜ 0.05 ),and the stimulate index was increased( P＜ 0.05 ) in the TGF-β2 antibody group; the expression of CD86,and MHC-Ⅱ was up-regulated (P＜0.05),the expression of dextran was down-regulated ( F =13.740,P =0.006 ),and the stimulate index was increased(P＜0.05) in the AH6809 group;the expression of MHC-Ⅱ was up-regulated and the stimulate index was increased with statistical difference in interaction(P＜0.05 ) in the AH6809+TGF-β2 antibody group.Compared with the control group,the expression of CD80 and CD86,and the stimulate index was still lower(P＜0.05 ).Conclusions TGF-β2 and PGE2 contribute to the inhibitory effects on DCs maturation mediated by murine CSCs in vitro and further have additive effect on the immunosuppression of DCs.

Objective To identify changes in the occurrence of endemic fluorosis in order to provide scientific basis for making countermeasures. Methods Five villages from 14 counties of mild, moderate and severe fluorosis affected areas were selected by stratified cluster sampling every year in the whole province during 2006 - 2010. Water and urinary fluorine were determined by ion selective electrode method(GB/T 8538-1995); dental fluorosis of children 8-12 years old was diagnosed with Dean method; skeletal fluorosis was diagnosed according to "clinical indexing standards of endemic skeletal fluorosis "(GB 16396-1996), between 2006 and 2008, and "clinical diagnosis standard of endemic skeletal fluorosis"(WS 192-2008) between 2009 and 2010. Results A total of 25 diseased villages were surveyed, 14 with water sources changed, covered a resident population of 8005 people, beneficiary population 7154, and accounting for 89.37% of the resident population; not changed villages 11. In accordance with the "State drinking water health standards", in the 14 changed villages the fluoride in drinking water was qualified (≤ 1.20 mg/L), there were 3 schools whose water fluorine content exceeded the standard; among the 11 villages that did not change water sources 7 drinking water samples fluorine content exceeded the standard. Of the 8 to 12 years old children in villages with changed water sources, 363 of them were checked and 142 dental fluorosis were found, the detection rate of dental fluorosis was 39.12% (142/363); in villages with water sources not changed, 303 children were checked, the detection rate of dental fluorosis was 43.89%(133/303). Of sixteen and elder adults in water source changed villages, 6424 people were checked and 403 skeletal fluorosis were found, skeletal fluorosis detection rate was 6.27% (403/6424); 3572 people were checked in not changed villages, the detection rate of skeletal fluorosis was 13.89%(496/3572). In water sources changed areas, geometric mean of urinary fluoride was in the normal reference value(WS/T 256-2005, 1.40 mg/L)or less. Conclusions Endemic fluorosis is decreased in water improved areas, but in unimproved areas the disease is still severe, and control of endemic fluorosis is still an arduous task.

Objective To evaluate the value of a CT ribs unfolding algorithm in the diagnosis of rib fracture. Methods Retrospective analysis of 180 patients who suffered chest trauma to do the chest or/and abdominal CT examination, and they also had the surgical or CT referral information. The images of these patients were postprocessed by software(Bone Reading software)and hand-drawn method(multi-point hand-painted CPR method). The rib fracture was observed and the time of reading was record. The diagnosis of fractures was confirmed by follow-up review or surgery. The fractures diagnosis sensitivity of the two post-treatment methods were measured, and the McNemar test was used to compare the difference between the software method and the hand-drawn method. Results Eight patients were excluded due to program failure, 172 cases were included in the study. Of the 172 patients, 63 patients suffered 259 fractures(178 ribs). The sensitivity of the software group was 91.7％(475/518), which was higher than that of the hand-painted group(86.3％, 447/518), the difference was statistically significant(P=0.005). The time of reading were (30.3 ± 3.3)and(173.2 ± 4.5)s, respectively, and the difference had statistically significant(P=0.001). Conclusion Compared to the traditional CPR method, the bone reading technique was used in patients with rib fractures during thoracic CT postprocessing can shorten the reading time and increase the sensitivity of the diagnosis.

BACKGROUNDChromosomal aberrations are the major cause of pre- and post-implantation embryo wastage and some studies suggest that half of all human conceptions have a chromosomal abnormality. A chromosomal aberration in human sperms is also one of the causes of failure of in vitro fertilization. This study was designed to ascertain whether chromosomal aneuploidy in spermatozoa is a risk factor for male infertility.

METHODSTwelve infertile men were divided into two groups: 10 with oligoasthenoteratozoospermia (OAT, Group A) and two with a normal semen analysis (Group B). Two normal healthy sperm donors acted as controls (Group C). We used fluorescence in situ hybridization (FISH) and probes for chromosomes X, Y and 18 to determine the frequency of aneuploidy.

RESULTSThe frequencies of spermatozoa disomy for chromosomes X, Y and 18 were 0.30% and 0.30%, respectively, in Group B. The percentages were not significantly different from those of Group C (0.15% and 0.16%). The frequencies of nullisomy for chromosomes X, Y and 18 were 0.15% and 0 for Group B, and 0 and 0.15% for Group C (P > 0.05). In Group A, the incidences of disomy were 1.13% and 0.96% and the frequencies of nullisomy were 1.13% and 1.60%. In these three groups, the incidences of diploidy were 0.60%, 1.00%, and 0.30%, respectively. Both the frequencies of disomic and nullisomic spermatozoa for chromosomes X, Y, and 18 and of diploid spermatozoa were significantly higher in Group A than in Groups B and C. The estimated total aneuploidy rates in the sperm from the three groups were 42.44%, 6.05%, and 2.59%, respectively.

CONCLUSIONThese results indicate that chromosomal aneuploidy in spermatozoa may be a risk factor for infertility.

Adult ; Aneuploidy ; Chromosomes, Human, Pair 18 ; Chromosomes, Human, X ; Chromosomes, Human, Y ; Humans ; In Situ Hybridization, Fluorescence ; Infertility, Male ; etiology ; genetics ; Male ; Risk Factors

By reverse transcription-polymerase chain reaction (RT-PCR), an open reading frame of pathogenesis-related protein 1 (PR1) was isolated from Panax notoginseng and named as PnPR1. Molecular and bioinformatic analyses of PnPR1 revealed that an open reading frame of 501 bp was predicted to encode a 166-amino acid protein with a deduced molecular mass of 18.1 kD. Homology analysis showed that the deduced amino acid sequence of PR1 protein of Panax notoginseng had a high similarity with other higher plants had the same conservative structure domain of cysteine-rich secretory protein (CAP). The recombinant expressed plasmid pET28a(+)-PnPR1 was expressed in Escherichia coli BL21. The expression conditions were optimized by induction at different times, different temperatures, different IPTG concentrations and different giving times. The optimum expression condition was 0.4 mmol.L-1 IPTG at 28 degrees C for 20 h. The successful expression of PnPR1 provides some basis for protein purification and preparation of the monoclonal antibody.
Amino Acid Sequence ; Cloning, Molecular ; Escherichia coli ; metabolism ; Molecular Weight ; Open Reading Frames ; genetics ; Panax notoginseng ; chemistry ; Phylogeny ; Plant Proteins ; genetics ; metabolism ; Plants, Medicinal ; chemistry ; Reverse Transcriptase Polymerase Chain Reaction ; Sequence Alignment

Now severe oligoasthenoteratozoospermic (OAT) patients could have offsprings because of the development of technique of intracytoplasmic sperm injection. But some researchers found these patients have increasing frequency of the aneuploid on the chromosome in their sperm. If the spermatozoa with chromosomal aneuploid were fertilized, it would be resulted in a higher rate of recurrent abortion, fetal abnormal and dead birth, so the analysis of the number of sperm chromosome will play an important role in detection on infertile men. There are many new development in the chromosomal analysis of human sperm using multi-FISH, now we have a review on them.
Aneuploidy ; Chromosomes ; Humans ; In Situ Hybridization, Fluorescence ; methods ; Infertility, Male ; genetics ; Male ; Spermatozoa ; physiology

BACKGROUNDAtherosclerosis is an important cardiovascular disease, becoming a major and increasing health problem in developed countries. However, the possible underlying mechanisms were not completely clear. In 2009, our research group first discovered that hydrogen sulfide (H(2)S) as a novel gastrotransmitter played an important anti-atherosclerotic role. The study was designed to examine the regulatory effect of hydrogen sulfide (H(2)S) on endoplasmic reticulum stress (ERS) in apolipoprotein E knockout (apoE(-/-)) mice fed a Western type diet.

METHODSC57BL/6 mice and homozygous apoE(-/-) mice were fed a Western type diet. C57BL/6 mice were injected intraperitoneally with normal saline (5 ml/kg per day) as control group. The apoE(-/-) mice were treated with the same dose of normal saline as the apoE(-/-) group, injected intraperitoneally with sodium hydrosulfide (NaHS, an H(2)S donor, 56 µmol/kg per day) as the apoE(-/-) + NaHS group and injected intraperitoneally with DL-propargylglycine (PPG, a cystathionine-γ-lyase inhibitor, 50 mg/kg, per day) as the apoE(-/-) + PPG group. After 10 weeks, the mice were sacrificed and the plasma lipids were detected. Sections of aortic root from these animals were examined for atherosclerotic lesions by HE and oil red O staining. The aortic ultrastructure and microstructure were analyzed with the help of light and electronic microscope. Glucose-regulated protein 78 (GRP78), caspase-12, copper-andzinc-containing superoxide dismutase (Cu/ZnSOD) and Mn-containing superoxide dismutase (MnSOD) protein expression in aortic tissues were detected with immunohistochemistry. The level of intracellular reactive oxygen species (ROS) were measured by using a commercial assay kit.

RESULTSCompared with control mice, apoE(-/-) mice showed increased plasma levels of total cholesterol (TC), triglyceride (TG) and low density lipoprotein (LDL), decreased high density lipoprotein (HDL), increased aortic plaque size, destroyed ultra-structure of aortic tissue, and increased expression of GRP78 and caspase-12 proteins. Compared with apoE(-/-) mice, H(2)S donor-treated apoE(-/-) mice showed a decreased plasma LDL level, lessened plaque necrosis and attenuated aortic ultra-structural disorder. H(2)S donor-treatment induced GRP78 expression but suppressed caspase-12 expression in aortic lesions. However, compared with apoE(-/-) mice, PPG treated apoE(-/-) mice showed enlarged plaque size, more severe ultrastructural disorder of the aortic tissue and reduced GRP78 staining in aortic lesions. The plasma lipids and the staining of caspase-12 in apoE(-/-) + PPG rats did not significantly differ from those in the apoE-/-mice. Consistently, H(2)S induced SOD expression, accompanied by a reduced level of ROS.

CONCLUSIONH(2)S plays a regulatory role in aortic ERS and reduces atherosclerotic lesions in apoE(-/-) mice fed with a Western type diet.

Animals ; Apolipoproteins E ; genetics ; metabolism ; Atherosclerosis ; blood ; Body Weight ; drug effects ; Cholesterol ; blood ; Endoplasmic Reticulum Stress ; drug effects ; Hydrogen Sulfide ; metabolism ; Lipoproteins, HDL ; blood ; Lipoproteins, LDL ; blood ; Male ; Mice ; Mice, Inbred C57BL ; Mice, Knockout ; Microscopy, Electron, Transmission ; Reactive Oxygen Species ; metabolism ; Sulfides ; pharmacology ; therapeutic use ; Triglycerides ; blood

Objective: To observe the effects of liraglutide on angiogenesis and myocardium protection in acute myocardial infarction (AMI) rats with its mechanisms. Methods: Rat's AMI model was established by left anterior descending of coronary ligation. AMI rats were randomly divided into 3 groups: Control group, the rats received subcutaneous injection of normal saline, Low dose (LS) group and High dose (HS) group, rats received subcutaneous injection of liraglutide 70μg/(kg?d) and 140μg/(kg?d) respectively; in addition, Sham operation group, rats received normal saline. n=6 in each group, all animals were treated for 2 weeks. 4 weeks later, cardiac structure and function were assessed by echocardiography, morphological changes of myocardium were observed by HE staining, collagen volume fraction (CVF) was calculated by Masson staining, myocardial microvessel density (MVD) and protein expression of vascular endothelial growth factor (VEGF) in marginal zone of infracted region were detected by immunohistochemistry, VEGF protein level was examined by Western blot analysis. Results: Compared with Sham operation group, Control group showed decreased LVEF, LVFS and increased LVEDd, LVESd, CVF, all P<0.01; while MVD and VEGF protein level were similar between 2 groups, P>0.05. Compared with Control group, LS group and HS group had obviously increased LVEF, LVFS, P<0.01 and decreased LVEDd, LVESd, P<0.05, obviously decreased CVF, P<0.01; obviously elevated MVD and VEGF protein level, P<0.01. Compared with LS group, HS group presented obviously increased LVEF, LVFS, P<0.01 and decreased LVEDd, LVESd, CVF, P<0.05; elevated MVD and VEGF protein level, P<0.01 or P<0.05. Conclusion: Liraglutide could improve angiogenesis in AMI rats which might be related to increased VEGF expression and reduced collagen deposition; therefore improve left ventricular systolic function for cardiac protection. The effect had certain relationship to liraglutide dosage.

OBJECTIVETo evaluate the effect and results of short and medium periods of follow-up of percutaneous balloon pulmonary valvuloplasty for critical pulmonary stenosis of neonates and infants under 6 months of age.

METHODSBetween January 2002 and December 2008, 34 consecutive patients aged from 13 to 175 days with critical pulmonary valvular stenosis underwent percutaneous balloon valvuloplasty. Patients records, catheterization data, angiograms and echocardiograms were reviewed. Patients were followed up for 6 months to 4 years (mean 25.5 months) by means of clinical examination and Doppler echocardiography.

RESULTSThe pulmonary valvuloplasty was accomplished in 32 (94%) of 34 attempts. Immediately after dilation, right ventricular systolic pressure (RVSP) decreased from (96 ± 28) mm Hg (1 mm Hg = 0.133 kPa) (49 ± 20) mm Hg (P < 0.01), the transvalvular peak to peak systolic gradient (ΔP) decreased from (89 ± 25) mm Hg to (25 ± 12) mm Hg (P < 0.01), and the right ventricular/aortic systolic pressure ratio decreased from 1.2 ± 0.5 to 0.7 ± 0.3 (P < 0.01). One patient died because of cardiac tamponade following rupture of the pulmonary valve annulus, 2 patients developed pericardial effusion, 3 patients had infundibular spasm, 3 patients had a pre-dilation by small balloon and 1 patient had weakened femoral artery pollex. After a follow up period of 6 months to 4 years 3 of 31 patients lost to follow-up. Repeat valvuloplasty was performed in 5 patients (3 neonates), no patient required surgery, and the other 23 patients did not undergo further intervention, a mean peak systolic Doppler gradient of (20 ± 13) mm Hg was found and no significant pulmonary regurgitation was seen.

CONCLUSIONSPercutaneous balloon pulmonary valvuloplasty was effective and safe for the treatment of critical pulmonary stenosis of neonates and infants under 6 months of age with good short and medium term results.

Catheterization ; Female ; Follow-Up Studies ; Humans ; Infant ; Infant, Newborn ; Lost to Follow-Up ; Male ; Pulmonary Valve ; Pulmonary Valve Stenosis ; therapy ; Treatment Outcome