The discovery of limbal stem cells is the key advancement in ophthalmology in the 20th century.Many clinical researches have proved that limbal stem cells transplantation is an effective method to treat severe ocular surface diseases.However, some important issues associated with the identification,isolation,standardized culture in vitro and differentiation after transplantation are still not clear and need to be paid more attention.Strengthening the underlaying research in this field will play important role in treating corneal blindness.

Child ; Choristoma ; Humans ; Male ; Neck ; pathology ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; Thymus Gland ; pathology ; Thymus Neoplasms

Objective To detect the gene expression of Pax6 in retinoblastoma (Rb).Design Experimental study.Participants Six cases of fresh Rb tumors and two established Rb cell lines.Six cases of retina tissues as normal control group.Methods Pax6 gene ex- pression was assessed in two established Rb cell lines,six primary tumors and six cases of retina tissues using the reverse transcriptase polymerase chain reaction (RT-PCR) and Western blot.Main Outcome Measures Expression of Pax6 gene in fresh retinoblastoma tis- sue and cell lines.Results The mRNA of Pax6 gene was positive in all two established Rb cell lines and the six primary Rb tumors using RT-PCR.The protein of Pax6 gene was also positive in Western blot.Pax6 mRNA and protein expression in the Rb tumors were increased compared with normal control group (P

AIMTo explore effects of Ginsenosides (Rb1, Rg1) on the expression of Bcl-2, Bax in the serum of kidney ischemia/reperfusion inducing apoptosis of HK-2 cells.

METHODSThe serum of rabbits with renal ischemia/reperfusion (SIR) and the control serum of rabbits (SC) were acquired and cultured with HK-2 cells. Detected apoptosis with TUNEL assay. The experiment was designed as: control group,ischemia/ reperfusion group, Rb1 blocking group and Rg1 blocking group. To detect the expression of Bcl-2, Bax with immunocytochemistry after 24 hours' cultured.

RESULTSThe expression of Bax in Rb1 blocking group and Rg1 blocking group were significantly decreased (P < 0.01), the ratios of Bcl-2/Bax were increased as compared with ischemia/reperfusion group.

CONCLUSIONRb1 and Rg1 have protective effects on apoptosis of HK-2 cells induced by serum of kidney ischemia/reperfusion.

Animals ; Apoptosis ; drug effects ; Cell Line ; Female ; Ginsenosides ; pharmacology ; Ischemia ; blood ; Kidney ; blood supply ; Kidney Tubules, Proximal ; cytology ; Male ; Proto-Oncogene Proteins c-bcl-2 ; genetics ; metabolism ; Rabbits ; Reperfusion Injury ; blood ; Serum ; physiology ; bcl-2-Associated X Protein ; genetics ; metabolism

Objective To investigate application of modified abdominal closure technique by an all layer in ventro-pelvic part operation and evaluate its value and significance. Methods 3200 cases with ventre-pelvic part operation between May 2002 and Aug 2007, were subjected to single layer closure with non absorbable suture material. The clinic data and some results of follow-up were retrospectively analyzed. Results Operative incisions of 1780 (55.6%) cases were in epigastric zone and their rate of primary healing was 98.5%, others (44.3%) in hypogastric zone (including pelvic cavity) and rate of primary healing was 98.2% (P>0.05). Rate of primary healing in older age-group was 97.9% and control group 98.8%, and primary healing of group diabetes 97.4%, control group 98.2% (P > 0.05). Average time of abdmenal closure was only 11±4 min. Primary complications included dehiscence of wound (0.5%), infection (1.4%) and incisional hernia (0.2%). Follow-up (66%) was performed at 30 days, 3 and 6 months, and at 1, 2 and 3 years. Conclusion It is concluded that closure of an abdominal incision can be effected by a multifilament interrupted absorbable sutures without an increased risk of wound dehiscence or incisional hernia, meanwhile economic and fast. It is the optimal method of abdominal closure and can be generalized.

objective To explore the role of cytokines and dendritic cells (DCs) in rat high-risk corneal allograft survival pro- longed by superantigen Staphylococcal Enterotoxin B (SEB) and to compare the different effects between SEB and glucocorticoid. Design Experimental study.Participants Fisher 344 and Lewis rats.Methods The Fisher 344 and Lewis inbred rats were used for high-risk penetrating keratoplasty model.All of the Lewis rat recipients were divided into three groups by blinded fashion.GroupⅠand GroupⅡrats were injected intraperitoneally with 0.2ml saline buffer or SEB (75?g/ml) respectively at 4-day intervals on three occasions before transplantation.GroupⅢrats were injected subconjunctivally with 0.1ml dexamethasone (1mg/ml) daily from the first day after surgery for 2 weeks.The allograft survival was examined under slit-lamp.The concentration of interleukin IL-1?,IL-2,IL-4,IL-5,IL-6, IL-10,TNF-?in rat aqueous humor and peripheral blood were measured by liquichip and the cytokine and CD11c,CD80,MHC-Ⅱex- pression in corneal grafts were examined by immunohistochemestry staining.Main Octcome Measures Mean survival time of the allo- grafts,the level of cytokines in aqueous humor,peripheral blood and corneal grafts.Results Compared with group control,the grafts mean survival time was delayed about 3.8d in group SEB (P=0.00) and 7.1d in group dexamethasone(P=0.01).But there was no signifi- cant difference between group SEB and dexamethone (P=0.26).Liquichip test showed that the level of IL-1?in aqueous humor was re- duced and IL-4,IL-6 and IFN-?,ascended.Only IFN-?,and IL-6 could be found in peripheral blood,and the changing shift of them was similar to that in aqueous humor.The immunohistochemistry staining showed that the expression of IL-2 in rat corneal grafts was signif- icantly decreased but IL-4,IL-6 and IL-10 elevated.The expression of DCs in group SEB was similar to that in group control,which was elevated after keratoplasty,but the phenotype of DCs was not the same.There were predominantly mature DCs in group control while immature DCs in group SEB.Conclusions The immunological inhibiting effect of SEB is same to glucocorticoid,but the mecha- nism is different.SEB can modulate immune response,which might induce immune inhibition via the local production of cytokines and effect on DCs maturation involving corneal graft rejection prevention.

Objective To introduce the experience and analyze the clinical outcome of applica- tion of reinforced bar-ring hybrid external fixator combined with limited internal fixation for complex frac- turcs of tibia plateau in patients.Methods From Aug.2002 to Feb.2005,21 patients with complex fractures of tibial plateau were treated with reinforced bar-ring hybrid external fixatar combined with lim- ited internal fixation.According to Schatzker classification,there were 13 patients with typeⅤfracture and 8 with typeⅥfracture.Voluntary and non-weight beating knee joint motion to partially weight bear- ing was taken one week after operation.Progressive loading was taken continuously until there was clinical evidence of fracture stability and then full weight bearing was done.Results The mean follow-up peri- od was 15.9 months (10-24 months).All the fractures got united and the mean union time was 4.2 months(3-8 months).All the injured knees could extend to less than 5 degrees and flex to more than 90 degrees after rehabilitation.Evaluated by Merchant score system for knee joint,81% of patients achieved excellent healing (7 cases) and good healing (10 cases).Conclusion Operation by using reinforced bar-ring hybrid external fixator combined with limited internal fixation can reduce complications and avoid stiffness of the knee joint.It is an effective method for complex fractures of tibial plateau.

OBJECTIVETo study the characteristics of gene expression of adrenal cortical steroid synthetase and its regulatory factor in mice with H22 liver cancer of different patterns.

METHODSSyndromes revealed in mice with H22 tumor were differentiated by quantified four diagnostic methods and syndrome differentiation, and mice with commonly encountered patterns (A: evil-toxin accumulation pattern, B: qi-deficiency pattern, C: yang-qi deficiency pattern and D: qi-yin-yang deficiency pattern) were screened out for subjecting to the study. Two batches of GeneChip Mouse Exon 1.0 ST Array detection were performed in the selected mice for detecting the gene expressions of adrenal cortical steroid synthetase and its regulatory factor, with the analysis performed put stress on the differential expressions in mice of various syndrome patterns.

RESULTSData obtained from the two batches detection showed well repeatability, in which similar genes of high or low expression emerged. The adrenal cortical steroid synthetase genes, such as Cyp11a1, Star, Cyp11b2, Cyp21a1, Hsd3b and Hsd17b were highly expressed, with few difference among the four patterns. However, Cyp11a1 was down-regulated and Cyp1b2 up-regulated in all patterns; Hsd3b1 and Cyp21a1 down-regulated in pattern A and B, but up-regulated in pattern C and D. As for the expressions of the relative regulatory factors, Cyb5b and Wnt4 were down-regulated but Fdx1, Fdxr, Hsd11b1, Por, Agt and Nr 0b1 were up-regulated in all patterns; Nr5al down-regulated in pattern A but up-regulated in other three patterns; Nr4al and Nr4a2 up-regulated in pattern A and down-regulated in the others.

CONCLUSIONSThe adrenal cortical steroid synthetase genes are rather conservative and stable in mice bearing H22 liver cancer, part of the expression might be correlated to the condition of disease and essence of syndromes, embodying the differences among different patterns in the same disease.

Adrenal Cortex ; metabolism ; Animals ; Gene Expression ; Gene Expression Profiling ; Gene Expression Regulation, Neoplastic ; Liver Neoplasms ; diagnosis ; genetics ; metabolism ; Male ; Medicine, Chinese Traditional ; Mice ; Mice, Inbred Strains ; Oligonucleotide Array Sequence Analysis ; Oxidoreductases ; genetics ; metabolism ; Steroids ; biosynthesis ; Yang Deficiency ; Yin Deficiency

OBJECTIVETo explore the effects of mifepristone on the growth of human gastric cancer cell line MKN-45 and its possible mechanisms.

METHODSIn situ hybridization was used to detect the expression of progesterone receptor (PR) mRNA in MKN-45 cells. Proliferation, cell cycle distribution, and the expression of Bcl-xL and vascular endothelial growth factor (VEGF) of MKN-45 cells incubated with various concentrations of mifepristone (1, 5, 10, and 20 micromol/L) were analyzed using MTT reduction assay, flow cytometry, reverse transcription-polymerase chain reaction (RT-PCR), and enzyme-linked immunoabsorbent assay (ELISA), respectively. After transplantation of MKN-45 cells underneath the skin of athymic mice, mifepristone was administrated with the dose of 50 mg/(kg x d) for 6 weeks to evaluate the tumor growth. Apoptosis and the expression of proliferating cell nuclear antigen (PCNA) in xenografted tumors were detected using transmission electron microscopy and immunohistochemical staining, respectively.

RESULTSPR mRNA was highly expressed in cultured MKN-45 cell. Mifepristone dose-dependently inhibited the proliferation of MKN-45 cells, and the inhibitory rate was dramatically increased from 7.21% to 47.23%. The inhibitory effect was accompanied by a dose-dependent increase in the percentage of cells in G0/G1 phase, and with a concurrent decrease in the proportion of S- and G2M-phase cells and the proliferative index from 57.65% to 24.54%. Meanwhile, mifepristone down-regulated the expression of Bcl-xL and VEGF in a dose-dependent manner. In vivo, mifepristone effectively inhibited the growth of xenografted tumors in nude mice (55.14% for inhibitory rate), induced apoptosis, and down-regulated PCNA expression in gastric cancer.

CONCLUSIONMifepristone exerts significant growth inhibitory effects on PR-positive human MKN-45 gastric cancer cells via multiple mechanisms, and may be a beneficial agent against the tumor.

Animals ; Apoptosis ; drug effects ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Dose-Response Relationship, Drug ; Down-Regulation ; Humans ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Mifepristone ; pharmacology ; RNA, Messenger ; biosynthesis ; genetics ; Receptors, Progesterone ; biosynthesis ; genetics ; Stomach Neoplasms ; metabolism ; pathology

OBJECTIVETo study the effect of osthole (Ost) on adrenocortical function in Y1 mouse adrenocortical tumor cells.

METHODSY1 mouse adrenocortical tumor cells were taken as subjects in this experiment. In 10.0%, 1.0%, and 0.1% serum DMEM-F12 medium, Y1 cells were treated with 1, 10, 25, 50, 100, and 200 micromol/L Ost for 24 and 48 h. 0.1% Dimethyl Sulfoxide (DMSO) was taken as negative control group and 1 mmol/L (Bu) 2cAMP as positive control group. Cell growth morphology was observed under inverted microscope. Contents of corticosterone were tested by ELISA. Expression levels of steroids synthase such as Star, Cyp11a1, Cyp21a1, Hsd3b2, Cyp11b1, Cyp11b2, Cyp17a1, and Hsd17b3 mRNA were detected by Real time quantitative PCR (RT-qPCR).

RESULTSY1 cell proliferation was obviously inhibited by 100 and 200 micromol/L Ost, and its inhibitory effect was more significant in 0.1% serum medium. Compared with the negative control group, gene expressions of Star, Cyp11a1 , Cyp21a1, Hsd3b2, Cyp11b1, Cyp17a1, and Hsd17b3 were significantly enhanced in the posi- tive control group (P < 0.05). Y1 cell corticosterone levels significantly increased in 50 micromol/L Ost treatment group after 24-and 48-h intervention (P < 0.05). Contents of corticosterone increased more obviously in 25 and 50 +/- mol/L Ost treatment groups after 48-h intervention, as compared with 24-h intervention (P < 0.01). After 24-h intervention, expression levels of Star, Cyp21a1, and Hsd3b2 genes were significantly up-regulated in 25 and 50 lLmol/L Ost groups (P < 0.05). Star gene expression was further enhanced after 48-h intervention (P < 0.05). However, Ost showed no effect on Cyp11a1 (P > 0.05). Additionally, gene expressions of Cyp11b1 and Cyp17a1 were significantly enhanced by 10, 25, and 50 pLmolIL Ost after treatment for 24 and 48 h (P < 0.05). Ost showed no obvious effect on Cyp11b2 and Hsd17b3 expressions.

CONCLUSIONOst could regulate adrenal cortex function and promote corticosterone synthesis and secretion through strengthening gene expressions of steroidogenic enzymes.

Adrenal Cortex ; drug effects ; Adrenal Cortex Neoplasms ; pathology ; Animals ; Corticosterone ; biosynthesis ; Coumarins ; pharmacology ; Gene Expression ; Mice ; RNA, Messenger ; metabolism ; Tumor Cells, Cultured