AlM: To study the therapeutic effect of external-route microsurgery forrhegmatogenous retinal detachment.
METHODS: ln 55 patients ( 55 eyes ) with rhegmatogenous retinal detachment, drainage of subretinal fluid, examination of locating the holes, sclera cryotherapy, scleral buckling, and vitreous cavity injection of filtrated air were performed under surgical microscope.
RESULTS:The retinal reattachment occurred in 50 cases after the primary surgery. The final rate of reattachment was 91% during 6 - 12mo follow - up. The retinal reattachment occurred in 1 case ( recurrent retinal detachment) after the secondary surgery and in 4 cases ( recurrent retinal detachment ) after vitrectomy. The eyesight was improved with different degrees in 55 cases.CONCLUSlON: The external- route microsurgery for rhegmatogenous retinal detachment is simple, safe and effective.

Objective To observe the clinical effect of interferon on the treatment of virus keratitis. Meth-ods Review and analysis was made of 476 patients with virus keratitis who was treated with high concentration of an-ti-virus eyedrops and one million unit of α-2b interferon, the clinical safety and effect was evaluated. Result The total cure rate was 59. 1%, and the type from high to low is interstitial、endothelial、epithelial and the total cornea. The total recurrence rate is 23.5% ,and the type from high to low is epithelial,the total cornea,endothelial and inter-stitial. The incidence rate of the adverse effect is 10. 7%. Condusion Systemic administration of interferon has a direct anti-virus effect, and it can raise the cure rate of virus keratitis as well as decrease recurrencerate. One million unit of interferon has a high clinical safety and effect.

Objective To construct a eukaryotic expression plasmid of pcDNA 3.1-Ag85A-CD226, and to use it as DNA vaccine then further evaluate its immunogenicity through oral administration in a mouse model.Methods The CD226-PCR2.1-ToPo plasmid was used as the template to clone CD 226 gene by PCR.The CD226 gene was then inserted into pcDNA 3.1-Ag85A plasmid to construct the recombinant plas-mid of pcDNA3.1-Ag85A-CD226.After identified by restriction enzyme analysis and sequencing , the re-combinant plasmid was transfected into HEK 293 cells by using lipofection .The expression of Ag85A-CD226 gene in HEK293 cells was detected by RT-PCR, Western blot and indirect immunofluorescence assay .The purified recombinant plasmid was used to prepare the Ag 85A-CD226 DNA vaccine by liposomal encapsula-tion.The vaccine was administered intragastrically to mice .The activities of NK cells , the cytokine levels in the supernatants of spleen cell cultures and the mRNA level of cytokines in the intestines were evaluated to analyze the immunogenicity of Ag85A-CD226 DNA vaccine.Results The Ag85A-CD226 DNA vaccine was prepared successfully .The expression of Ag85A-CD226 fusion protein was detected in HEK293 cells.The activities of NK cells from mice vaccinated with Ag 85A-CD226 DNA vaccine were higher than those from other control groups (P<0.01).The level of TNF-α, IFN-γand IL-2 in the supernatants of spleen cell cul-tures and in the intestines were significantly up-regulated in comparison with other control groups ( P <0.01).The level of IL-4 in the supernatants of spleen cell cultures was down-regulated in the experimental group (P<0.01), but the level of IL-4 in intestines showed no significant difference among the five groups (P>0.05).Conclusion The Ag85A-CD226 DNA vaccine could significantly enhance Th1 type immune responses systemically and in the intestine as in comparison with those vaccinated with single dose of Ag 85A DNA vaccine or CD226 DNA vaccine.

To construct, express and purify fusion protein containing HBcAg and HBsAg preS1 epitope peptide for the purpose of investigating a novel HBV vaccine with both prophylactic and therapeutic functions. Using DNA recombinant technology, prokaryotic expression plasmid pBTcs1 expressing HBcAg and HBsAg pre-S1 epitope peptide fusion protein was constructed. After expressed in E.coli. HB101, the production BTcs1 was purified by sucrose density gradient ultracentrifugation and identified by SDS-PAGE, SEC, Western-blot and electron microscope. The results indicated that expression plasmid pBTcs1 was constructed successfully, and 20~25 mg purified BTcs1 fusion protein was obtained from 1L LB culture. Result of DOT-BLOT indicated that the distribution of BTcs1 was mainly in 30~50% sucrose, the purity of BTcs1 was greater than 95% by SDS-PAGE and SEC analysis. BTcs1 could probe with specific antibodies at 28 kDa by Western-blot, BTcs1 could also self assemble VLP by electron microscope analysis, its diameter was 30~34 nm approximately. The present study lay a foundation for further research functions and applications of BTcs1.

Objective：To explore the difference in creative thinking and the related factors between deaf children and normal children.Methods：Observation group(n=122)with the hearing disability students were selected from 4 special education schools.Control group(n=122)was come from 2 ordinary primary schools and 2 ordinary middle schools.The two groups were given both the New Creativity Test and the Combined Raven's Test.Results：(1)Deaf children got lower scores than normal children in verbal fluency[(7.76±0.75)vs.(12.98±0.59),P<0.001],verbal flexibility[(4.28±0.33)vs.(7.87±0.28),P<0.001],verbal originality [(7.16±0.89)vs.(11.35±0.72),P<0.001],figural flexibility[(9.69±0.35)vs.(11.10±0.31),P=0.003]and IQ[(101.05±1.196)vs.(105.01±1.102),P=0.030].Deaf children got higher scores than normal children in figural elaboration[(3.24±0.40)vs.(1.96±0.22),P=0.006].There was no significant difference in fluency and originality of figural task between the two groups.(2)Deaf children's scores of verbal fluency and verbal originality were positively correlated with their age(β=0.310,0.301;Ps<0.01).Deaf children's scores of verbal flexibility were positively correlated with length of bilingual education(β=0.308,P<0.001).Deaf children's scores of figural fluency,figural flexibility,figural originalityand figural elaboration were correlated positively with their age of sign language(β=0.321,0.308,0.228,0.456;Ps<0.05).Conclusions:(1)Deaf children are lower than normal children in verbal fluency,verbal flexibility,verbal originality,figural flexibility,and are higher in figural elaboration.There is no difference in figural fluency and originality between them.(2)Sign language is a major related factor to deaf children's figural creative thinking.

Objective:To prepare a kind of G418 resistance fibroblast feeder layers.Methods: In order to prepare a stock primary embryonic fibroblast (EMFI) cells, the pregnant Smad3ex8+/- mice containing heterozygous neo gene were killed 14 days after coitus to get the embryo. EMFI cells were tested by PCR to identify their gene types. Only one EMFI cell which contained heterozygous neo gene was chosen and treated with mitomycin C.The mitomycin C-treated EMFI feeders were stored at -70℃. Results: The EMFI feeder layers could support the growth of embryonic stem cell line TC-1 efficiently and could live in G418 resistance medium for 1-2 weeks.

OBJECTIVETo investigate the therapeutic effect of glucocorticoids on the acute pulmonary edema in rats induced by nitrogen dioxide (NO₂).

METHODSThirty SD female rats were randomly equally divided into 5 groups: normal control group, NO₂ exposed group, high-, middle- and low-dose of glucocorticoids treated group (6 rats per group). 6 rats in the normal control group were exposed to room air for 30 min, and the other rats to NO₂. 18 rats in the glucocorticoids group were treated with different doses of dexamethasone (6.0, 3.0, 1.0 mg/kg), while the rats in the NO₂ poisoning group were treated with normal saline (2.5 mg/kg). The lung wet/dry (W/D) weight ratio was calculated, and plasma atrial natriuretic peptide (ANP) levels, superoxide dismutase (SOD) activity from whole blood, plasma interleukin 6 (IL-6), interleukin 10 (IL-10), tumor necrosis factor-α (TNF-α) and Interferon-γ (IFN-γ) were detected by enzyme-linked immunosorbent assay (ELISA).

RESULTSThe lung W/D ratios were increased significantly in glucocorticoids treated group and NO₂-exposed group compared with normal control group (P < 0.05), while they were significantly reduced in glucocorticoids treated group as compared with NO₂-exposed group (P < 0.05). SOD activity in whole blood in glucocorticoids treated group and NO₂-exposed group was significantly lower than that of normal control group (P < 0.05), while it was no significant difference between that of glucocorticoids treated group and NO₂-exposed group (P > 0.05). Plasma ANP was significantly increased in NO₂-exposed group compared with normal control group (P < 0.05), while it was significantly decreased in glucocorticoids treated group compared with NO₂-exposed group (P > 0.05). Plasma TNF-α of high-, middle- and low-dose of glucocorticoids treated group [(27.04 ± 8.19), (40.10 ± 9.09), (39.76 ± 9.60) pg/ml] was decreased significantly as compared with NO₂-exposed group (68.55 ± 27.84 pg/ml) (P < 0.05). Plasma IL-6 in high- and middle-dose of glucocorticoids treated group [(15.97 ± 6.18), (19.69 ± 5.52) pg/ml] was significantly decreased as compared to NO₂-exposed group [(29.29 ± 9.31) pg/ml] (P < 0.05). Plasma IL-10 in high-, middle- and low-dose of glucocorticoids treated group [(23.24 ± 5.14), (27.78 ± 8.17), (33.29 ± 10.42) pg/ml] was significantly reduced compared with NO₂-exposed group [(44.38 ± 9.19) pg/ml] (P < 0.05). Plasma IFN-γ in high- and middle-dose of glucocorticoids treated group [(7.21 ± 4.55), (19.23 ± 4.35) pg/ml] was reduced compared with NO₂-exposed group [(30.83 ± 6.82) pg/ml] (P < 0.05).

CONCLUSIONHigh-, middle-, low-dose glucocorticoids all can improve the permeability of alveolar wall and capillary, and have nonspecific anti-inflammatory effects. The therapeutic effects on pulmonary edema are significant. High and middle dose of glucocorticoids treated group are more useful for decreased inflammatory factors.

Animals ; Disease Models, Animal ; Female ; Glucocorticoids ; therapeutic use ; Nitrogen Dioxide ; toxicity ; Pulmonary Edema ; chemically induced ; drug therapy ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo observe the effect of single herb pilose antler (PA) on the expression of Smad2 and Smad3 in the cartilage of osteoarthritis (OA) rats.

METHODSOne hundred 3-month old female healthy SD rats, (200 +/- 20) g, were recruited and routinely fed for 1 week. They were randomly divided into 5 groups, i.e., the low dose PA group, the high dose PA group, the normal saline control group, the model group, and the normal control group, 20 in each group. The model was prepared using classic Hulth method except the normal control group. After 6-week modeling, the model was confirmed successful by pathologic observation. PA at 0.021 g/100 g and 0.084 g/1 00 g was given by gastrogavage to rats in the low dose PA group and the high dose PA group respectively. Normal saline was administered to those in the normal saline control group. No treatment was given to rats in the normal control group and the model group. Bilateral knee cartilages were harvested at week 2,4, and 6. mRNA and protein expressions of Smad2 and Smad3 were detected by immunohistochemical assay, fluorescent quantitative PCR, and Western blot.

RESULTSOA model was successfully prepared by pathological observation. Results of immunohistochemical assay showed that Smad2 and Smad3 expressed extensively in the cartilage, and located inside the chondrocyte membrane. Compared with the model group, mRNA expression of Smad2 and Smad3 obviously increased in the low dose PA group and the high dose PA group at week 2, 4, and 6, showing statistical difference (P < 0.05). Compared with the same group at week 4 after gastrogavage, mRNA expression of Smad2 and Smad3 obviously decreased in the low dose PA group and the high dose PA group at week 6, showing statistical difference (P < 0.05). Compared with the model group, protein expression of Smad2 and Smad3 obviously increased in the chondrocytes of the low dose PA group and the high dose PA group at week 2 and 4, showing statistical difference (P < 0.01). Compared with the same group at week 2 after gastrogavage, protein expression of Smad2 and Smad3 obviously increased in the low dose PA group and the high dose PA group at week 4, showing statistical difference (P < 0.01). Compared with the same group at week 4 after gastrogavage, protein expression of Smad2 and Smad3 obviously decreased in the low dose PA group and the high dose PA group at week 6, showing statistical difference (P < 0.01).

CONCLUSIONS(1) The pilose antler could repair cartilages by regulating mRNA and protein expressions of Smad2 and Smad3. (2) Up-regulating mRNA and protein expressions of Smad2 and Smad3 might be one of important mechanisms for the pathogenesis of OA.

Animals ; Antlers ; chemistry ; Cartilage ; cytology ; metabolism ; Chondrocytes ; drug effects ; metabolism ; Female ; Medicine, Chinese Traditional ; Osteoarthritis ; drug therapy ; metabolism ; Rats ; Rats, Sprague-Dawley ; Smad2 Protein ; metabolism ; Smad3 Protein ; metabolism

Objective:To investigate the effects of interleukin-34(IL-34)on the expression of CC chemokine ligand 28 (CCL28) by fibroblast-like synoviocytes(FLS) in rheumatoid arthritis(RA) patients.Methods:FLS were isolated from 6 RA patients and stimulated with IL-34,IL-34 receptor antagonist /IL-34,inhibitors of signaling pathway/IL-34 in vitro respectively.CCL28 mRNA expression was measured by reverse transcription polymerase chain reaction(RT-PCR).The level of CCL28 in the supernatant of RA FLS culture was detected by enzyme linked immunosorbent assay(ELISA).Statistical analysis between groups was performed by t test.Results:Compared with unstimulated FLS,CCL28 expression was increased obviously in IL-34-stimulated group(P<0.05).The level of CCL28 was significantly decreased when anti-IL-34 antibody was added into IL-34-administrated RA FLS(P<0.05).While after adding of nuclear factor κB(NF-κB) and mitogen-activated protein kinase38(p38 MAPK) signaling pathway inhibitors into the cell culture system,CCL28 expression was remarkably reduced (P<0.05).Conclusion: The secretion of CCL28 by RA FLS can be promoted by the binding of IL-34 with its specific receptor via the activation of NF-κB and p38 MAPK signaling pathways,which suggests that CCL28 might be involved in the pathogenesis of RA.

Objective: To evaluate the clinical value of echocardiography in aortic valvuloplasty (AVP) in the low-age pediatric patients with congenital aortic valve stenosis. Methods: We retrospectively studied 39 low-age (at median of 23 months) patients with congenital aortic valve stenosis who received aortic valve repair in our hospital for their echocardiography information, and statistically analyzed the main indicator changes by 4 time points as pre-operation and 1 week, 1-3 months, 6-12 months after the operation respectively. Results: In our study, the bicuspid to tricuspid valve ratio was approximately at 5.5/1 and 2 patients died during peri-operative period. Compared with pre-operative time point, Doppler aortic valve peak velocity (Vmax ) and the mean aortic transvalvular pressure gradient (MPG) were reduced accordingly, for Vmax: (4.30 ± 0.73) m/s vs (2.65 ± 0.78) m/s, (2.93 ± 0.63) m/s, (3.01 ± 0.83) m/s,P<0.01, for MPG: (45.78 ± 15.19) mmHg vs (18.24 ± 10.08) mmHg, (21.01 ± 10.08) mmHg, (22.31 ± 13.41) mmHg. Compared with pre-operative time point, left ventricular ejection fractions (LVEF) were similar in 3 post-operative time points. Compared with 1 week post operative time point, left ventricular end-diastolic anteroposterior diameter (LVEDD) was increased at 6-12 months post-operative time point, the relative wall thickness (RWT) was decreased, bothP<0.05, and aorta valve regurgitation (AR) was increasedP<0.01. Pearson correlation analysis showed that aortic annulus (AA) inner diameter was positively related to LVEDD (r= 0.648,P<0.01), negatively related to Vmax (r= -0.205,P<0.05) and RWT was positively related to Vmax (r= 0.196,P<0.05). There were 6 patients with pre-operatively decreased LVEF, 1 of them died and the rest 5 with elevated LVEF at 6-12 months post-operative period,P<0.05. Conclusion: Echocardiograghy could be used as the ifrst choice of imaging method for diagnosing congenital aortic valve stenosis, it has the important role for in-operative monitoring and post-operative evaluation of AVP in relevant patients.