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Objective: To develvp a RP-HPLC method for the determination of flavonoids in fifteen kinds of flowers such as Iris lacteal pall, prunus persica and rosa chinensis., Method: The contents of quercetin, kaempferol and isorhamntin in fifteen kinds of flowers were extracted with methanol. The analysis was performed on a Kromasil C18 column (4.6 mm x250 mm, 5 microm) with methanol-0.1% phosphoric acid (50:50) as mobile phase., Result: The quercetin, kaempferol and isorhamntin were separated well, and the result shows that the content of quercetin in the Iris lactea Pall was the highest (1.536%), the contene of kaempferol in Persica persice was the highest (0.572%), and the content of isorhamntin in chrysamthemum morifolium was up to 0.290%., Conclusion: The contents of flavonoids in these flowers were by determined RP-HPLC for the first time and the method can be used for quantitative determination of flavonoids in the flowers.

The concept of "ischemic postconditioning" was first raised in 2002, and the following 5 year research shows that it can protect organs from reperfusion injury. Although the mechanism of ischemic postconditioning is similar to ischemic preconditioning in many ways, it still has its own characteristics. Reperfusion injury is an inevitable problem in organ transplantation. It may accelerate the function recovery of the transplants to lessen the reperfusion injury. So ischemic postconditioning may have a fine prospect in organ transplantation for its good controllability during reperfusion. This article is going to briefly introduce the distinct mechanisms of ischemic postconditioning to protect organs from reperfusion injury and approach the possibilities of its application in organ transplantation.

Objective: The aim of this study was to assess the protection of recombinant human erythropoietin (rhEPO) in light-induced injures in human retinal pigment epithelial (RPE) cells., Methods: It was a experimental study. Cultured human RPE cells were exposed to light of 8w 2000 +/- 500 Lux for 12 hours. The 3-(4,5-dimethylthiazole-2y1)-2,5-diphenyl tetrazolium bromide (MTT) cell viability assay were used to assess the effects of rhEPO in light-induced injury on human RPE cells. The effect of inhibiting apoptosis of rhEPO was detected by AnnexinV-fluorescein isothiocyanate/Propidium iodium labeling and flow cytometry. The enzyme linked immunosorbant assay (ELISA) and immunocytochemical staining were used to assess the expressions of caspase-3 treated by different doses of rhEPO in light-induced injury on human RPE cells and examine the protective mechanism of rhEPO by treatment with AG490 (the special inhibitor of jak2)., Results: There was a significant increase of inhibiting apoptosis in every rhEPO group, and cell viability was the highest in 40 U/ml rhEPO group, the value was 4.93 +/- 1.45/ml. The decrease in expression of caspase-3 was the most obvious in 40 U/ml rhEPO group, in which the value was 0.125 +/- 0.029 ng/ml. There was a significant increased effect on inhibiting apoptosis in every rhEPO group, and it was the most conspicuous in 40 U/ml rhEPO group. But these increased cell viability and effect on inhibiting apoptosis in rhEPO group were restrained by AG490, in which the value of apoptosis was 11.29 +/- 2.11/ml and the density of caspase-3 increased to 0.362 +/- 0.042 ng/ml., Conclusions: It is suggested that rhEPO can protect human RPE cells from the light-induced injures. Its protective mechanism is principally mediated by the EPO-EPOR pathway, which subsequently leads to jak2 activation.

Background: Bile leak remains a main complication in liver transplantation patients with poor biliary tract conditions, mainly caused by an insufficient blood supply or dysplasia of the biliary tract. Although Roux-en-Y modus operandi can be adopted, the risk of other complications of the biliary tract such as infection increases. Using pedicled greater omentum flaps to wrap the anastomotic stoma, which increases the biliary tract blood supply, may reduce the incidence of bile leak., Methods: Fourteen patients undergoing piggy-back liver transplantation and having poor biliary tract conditions were treated with pedicled greater omentum flaps to wrap the anastomotic stoma of the biliary tract. Their clinical data were analyzed retrospectively., Results: Of the 14 patients, only one (7.1%) had a mild bile leak on the 8th day post-operation and fully recovered after symptomatic treatment. The other patients had no biliary complications., Conclusions: Using pedicled greater omentum flaps to wrap the anastomotic stoma of the biliary tract is an effective way to prevent bile leak in liver transplantation patients, especially those with poor biliary tract conditions. However, experience with this surgical technique still needs to be further explored.

Objective: To investigate the expression of erythropoietin (EPO) in human fetal retinal pigment epithelium (hfRPE) cells exposed to oxidative stress induced by hydrogen peroxide (H(2)O(2)) and to study the mechanisms., Methods: The hfRPE cells were isolated, cultured and identified. The content of malondialdehyde (MDA) and the activity of superoxide dismutase (SOD) of the hfRPE cells were examined. The changes of level of mRNA and protein of EPO in hfRPE cells exposed to oxidative stress were measured by semi-quantitative RT-PCR and immunocytochemical staining techniques., Results: H(2)O(2) could increase the content of MDA and inhibit the activity of SOD in hfRPE cells. RT-PCR detected a significant increase of EPO mRNA level in cultured hfRPE cells exposed to oxidative stress. The level of EPO mRNA in the hfRPE cells reached a peak when exposed to 600 micromol/L H(2)O(2), then decreased after exposing to 800 micromol/L H(2)O(2). The immunocytochemical study detected that the changes of the level of EPO protein were similar to that of EPO mRNA., Conclusion: Oxidation stress by exposed to H(2)O(2) induces significant increase of EPO expression of hfRPE cells. Expression of EPO may be related to the survival and tolerance of hfRPE cells.

Two hybrid combinations were made with a resynthesized specially-long pod line in H218 Brassica napus L., namely H218 x Zhongyou 821, H218 x MSP334 and their six generations F(1), F(2), backcrosses B(1) and B(2) and their parents P(1) and P(2) were used to study the inheritance of 3 pod related traits, namely the full pod length, the pod body length and the pod beak length. The results were the followings: the three traits in the F(2) population of the two combinations had a continuous normal distribution, indicating that these traits were quantitative in nature and controlled by polygenes. The broad-sense heritabilities of the full pod length, pod body length and pod beak length of the two combinations were 65.89%-70.77%, 60.14%-63.38% and 26.36%-46.44%, respectively. The narrow-sense heritabilities were 44.01%-46.78%, 46.89%-47.38% and 18.08%-37.87%, respectively. The results from the two combinations consistently showed that full pod length, pod body length and pod beak length were controlled by 5, 6 and 2 genes respectively. The preliminary study on the gene effect demonstrated that the three traits in the two combinations all fit the additive-dominant model and the epistatic effect was significant. The additive effects and the dominant effects of the three traits in the two combinations were both significant. Of the full pod length and the pod body length, the dominant effect was more significant than the additive effect; and it is the same case for the pod beak length.

Objective: To study the clinical and pathological characteristics of choroidal metastatic carcinoma., Methods: The clinical data, pathological character, primary tumor origin and histological classification of 18 patients with choroidal metastatic carcinoma were analyzed retrospectively., Results: Most patients had severe visual impairment. Solid mass was seen in the posterior pole of the eyes in all 18 patients through ocular fundus examination, 8 cases had retinal detachment. B scan and CT examination found flat or irregular masses. MRI examination had been performed on 5 patients, high signal intensities on T1W and low signal intensities on T2W were found. Five patients were adenocarcinoma, 4 were squamous carcinoma and 3 were undifferentiated carcinoma through pathological examination. Primary tumor was lung carcinoma in 10 cases (55%) and breast carcinoma in 4 cases (22%)., Conclusions: Rapid decrease of visual acuity, flat neoplasm in ocular fundus and secondary retinal detachment are the main clinical characteristics of choroidal metastatic carcinoma. The most common primary tumor is lung carcinoma and the most common histopathological classification is adenocarcinoma. Imaging examination is helpful for the diagnosis of choroidal metastatic carcinoma.

Objective: To explore the effects of oxidative injury induced by hydrogen peroxide on human retinal pigment epithelial (RPE) cells., Methods: Cultured human RPE cells were treated by 600 micromol/L hydrogen peroxide (H2O2) for 1, 6, 12, 24 and 72 hours. Cell viability was assessed by the MTT assay. Apoptosis was assessed by Annexin V-fluorescein isothiocyanate/Propidium iodium (Annexin V-FITC/PI) staining. The expression of clusterin was assessed by Western blot., Results: (1) The treatment of RPE cells with 600 micromol/L H2O2 caused a time-dependent decrease of cellular viability. (2) Apoptosis was detected in cultured human RPE cells treated with 600 micromol/L H2O2 for 6 hours. The number of apoptotic cells reached a maximum at 24th hour after being exposed to 600 micromol/L H2O2 (P < 0.05). (3) Western blot showed the expression of clusterin protein was demonstrated in 6 hours exposure to 600 micromol/L H2O2, a significantly increasing of clusterin expression was observed overtime (P < 0.01). Thereafter the expression of clusterin protein decreased at 24th hour after being exposed to 600 micromol/L H2O2. At 72nd hour, the expression of clusterin protein was quite weak., Conclusion: Hydrogen peroxide can inhibits RPE proliferation and induces apoptosis and aging gene expression;the result suggest that accumulative oxidative injury induced by hydrogen peroxide in RPE in vitro may be similar to the aging changes in vivo.

Objective: To explore the technique of induction of polyploidy in Salvia bowleyana by colchicine treatment., Method: The three kinds of explant of bud, leaf and calli were induced by colchicine treatment., Result: The induction effects were better when the calli was treated by colchicines (15 mg x L(-1)) and the leaf was pre-cultured for one week. The doubling rate was 33.33%, while the majority were wholy doubled plants, and the leaves were thicker and broader, the color was darker, the root was thicker and the stoma size was obviously bigger than the diploid plants. The number of chromosome were 8 to 64. Isoenzyme analysis showed that the enzyme activities between the polyploid and the diploid plants were quite different., Conclusion: Induction of polyploidy by colchicine treatment is efficacious. The part of the doubled plants were identified as homologmous tetraploids.

The intergeneric hybrid from a cross between Brassica. Chinensis and Crambe abyssinica was observed with 2n=55 chromosomes in the original progenies. After several generations of in-vitro propagation by tissue culture, the chromosomes of the intergeneric hybrid were remarkably reduced, varying from 25 to 28, averaged at 26.In meiosis of the PMC of the hybrid, the average configuration of chromosome pairing was 0.06 III + 11.26 II + 3.80 I. The number of bivalents varied from 8 to 13. The majority of PMC cells showed 10 II, 11 II and 12 II bivalents with frequencies of 24.58%, 23.91% and 30.98% respectively. The number of univalents varied from 0 to 8. The reduction of chromosomes in the hybrid and the high numbers of bivalents were possibly due to the chromosome of Crambe abyssinica eliminating and the genome of Brassica. Chinensis doubling in the hybrid cells. Triade cells, chromosome lagging, and chromosome bridges were observed in anaphase II.

Objective: To study the protective roles of hypoxic preconditioning in light induced retinal injury in a mice model and investigate the possible mechanism of related gene regulation., Methods: 54 BALB/c mice were randomly divided into simple light exposure group (SL), hypoxic pretreatment group (HP) and control group (CON). The mice of SL and HP were continually exposed to light for 3 h, which built model of light-induced damage. Morphologic changes of photoreceptor cells in different group were examined by light microscope and the apoptosis was detected by TdT-mediated dUTP nick end labeling. Different expressions of c-fos and caspase-1 gene were examined by immunohistochemical staining., Results: In group SL, the photic injury appeared very obviously and early. Different changes appeared in the outer nuclear layer after light exposure. Photic injury was aggravated following the increased light exposure. Positive staining of c-fos and caspase-1 could be seen in the outer nuclear layer. In group HP, the changes of retinal morphology appeared slightly and lately. Compared with group SL, caspsae-1 expression was decreased obviously, while no difference was seen in the expression of c-fos. The retinal structures was normal in the mice of control group and c-fos and caspase-1 were stained negative., Conclusion: Hypoxic preconditioning has neuroprotective effect on photoreceptor cells by inhibiting the expression of apoptosis-related genes in photic injured mice model. caspase-1 may be involved in the protective mechanism.

Background: Ischemia/reperfusion is the main cause of hepatic damage in liver transplantation. Immediate early genes (IEGs) encode proteins can regulate expression of cellular response genes after injury, and is associated with tissue repair and cell apoptosis. The purpose of this research was to investigate the effects of preconditioning on expression of immediate early genes c-fos and c-jun following hepatic ischemia/reperfusion (IR) and its roles in cellular regeneration and apoptosis., Methods: Ninety-six Wistar rats were randomly divided into IR group and hepatic ischemic preconditioning (IPC) group, and each group was further divided into eight sub-groups (n=6). The model of partial liver ischemia/reperfusion was used. The rats were subjected to 60-minute liver ischemia, preceded by 10-minute preconditioning. After 0-, 0.5-, 1-, 2-, 4-, 8-, 12-, 24-hour reperfusion, the serum and liver tissue in each group were collected to detect the level of serum ALT/AST, liver histopathology, expression of c-fos, and c-jun mRNA. Flow cytometer was used to detect Ki67 and Sub-G1 as the quantity indicators of cell regeneration and apoptosis respectively., Results: Compared with IR group, IPC group showed a significantly lower ALT/AST level in 0.5-hour sub-group to 8-hour sub-group (P<0.05). Ki67 elevated significantly at 0.5, 1, 2 hours, but decreased significantly at 24 hours (P<0.05). Ap index decreased significantly after 1-hour reperfusion (P<0.05). Expressions of c-fos and c-jun mRNA were low, especially c-jun at 0.5, 1 and 2 hours after reperfusion., Conclusion: Ischemic preconditioning can protect liver cells against ischemia/reperfusion injury, and this protective effect may be related to influence transcription levels of c-fos and c-jun.

Objective: To study whether recombinant human erythropoietin can pass through mice blood-retina barrier and the protective role in light-induced damage in retina., Methods: After the injection of rHEPO, the content of rHEPO in 24 BALB/c mice retina was examined by enzyme linked immunosorbent assay (ELISA). 24 BALB/c mice were used to establish a light-induced damaged model, the difference of retina in rHEPO group and control group was compared using light microscope and TdT-mediated dUTP nick end labeling (TUNEL)., Results: The amount of retinal rHEPO in four deferent time points was (0.68 +/- 0.24) mU, (1.87 +/- 0.37) mU, (0.96 +/- 0.24) mU, (0.47 +/- 0.13) mU in 100 microg retinal total protein respectively by ELISA assay, there were statistical significances among groups. The density of rHEPO in the retina reached its peak at 4th hour after injection. Histology analysis: rHEPO group, at the 12th hour after light exposure the inner segment became condensed and disorganized. At the 36th hour the retina disorganized and vesiculated were seen in outer segments. At the 72nd hour the inner and outer segments were damaged more seriously and the outer nuclear layer became thinner and denser. On the 7th day, the retinal outer nuclear layer became thinner and condenses. rHEPO group showed a minimal damage in every time points but outer nuclear layer disorganized and vesiculated in inner and outer segments. No obvious changes in retinal thickness. The apoptotic cells were detected by TUNEL. At the 12th hour after light exposure, there were the apoptotic cells in outer nuclear layer near outer plexiform layer. At 36th hour the numbers of apoptotic cells were increased, however at the 72nd it was decreased obviously, only a few scattering apoptotic cells were revealed in the outer nuclear layer. Numbers of apoptotic cells between the rHEPO group and control group in outer nuclear layer were statistical significance (P < 0.01)., Conclusions: rHEPO can pass through the mice blood-retina barrier and rHEPO has neuroprotective effect on mice retina. rHEPO may be used to treat degenerative retinal diseases.

Objective: To investigate the expression of cluster of differentiation 44 variant 6 (CD44V6) and fibronectin (FN) in lacrimal gland tumors., Methods: Strept Avidin-Biotin Complex (SABC) immunohistochemistry method was used to study the expression of CD44V6 and FN in 49 cases of lacrimal gland tumors., Results: In benign lacrimal gland tumors, the CD44V6 and FN positive rates were 30.8% (8/26) and 80.8% (21/26) respectively. In malignant lacrimal gland tumors, the CD44V6 and FN positive rates were 60.9% (14/23) and 34.8% (8/23). The positive expression rates of CD44V6 and FN in malignance were significantly different from benign and control tissue. There is also a significant difference in the expression of CD44V6 and FN in the cases of recurrence and non recurrence (P <0.05)., Conclusions: The expression of CD44V6 and FN was correlated with the progress and recurrence of lacrimal gland tumors. The positive staining of CD44V6 and FN was useful for differentiating malignance from benignity and evaluating the progress and recurrence.

Background: Basic fibroblast growth factor (bFGF) plays important roles in retina degeneration, light injury, mechanical injury, especially in retina ischemia-reperfusion injury (RIRI). This study was to investigate the therapeutical effect of bFGF on RIRI and its mechanisms., Methods: Experimental RIRI was induced by increasing intraocular pressure (IOP) in the eyes of 48 rats. These rats were divided into normal control, ischemia-reperfusion and bFGF-treated groups. Histological and ultrastructural changes of in the retina of different groups were observed, and the number of retinal ganglion cells (RGCs) was quantitatively analyzed under microscopy. Apoptotic cells were detected using the TdT-dUTP terminal nick-end labeling (TUNEL) method. The expression of caspase-3 was determined by streptavidin peroxidase (SP) immunohistochemistry. Atomic absorption spectrum method was used to evaluate the intracellular calcium changes., Results: At the early stage of retinal ischemia-reperfusion injury, retina edema in the treated group was significantly eliminated compared with the untreated ischemic animals. RGCs in the bFGF-treated group was more than those in the untreated ischemic group during the post-reperfusion stages. In ischemic group, apoptotic cells could be found at 6th hour after reperfusion and reached the peak at 24 hours. At 72nd hour no apoptotic cells could be found.The changes in caspase-3 expression had a similar manner. The intracellular calcium of rat retina began to increase at 1st hour, reached the peak at 24 hours, and began to decrease at 72 hours. The change of the three markers in the treatment group showed a similar pattern, but they were all relatively less obvious., Conclusion: Apoptosis may play a vital role in RIRI. bFGF may has therapeutical effects on RIRI by inhibiting the increase of intracellular calcium and caspase-3 expression.

Objective: To investigate the therapeutical effect of basic fibroblast growth factor (bFGF) on retina ischemia/reperfusion injury., Method: Experimental retinal ischemia/reperfusion injury was induced by increasing intraocular pressure of rats eyes. 48 rats were divided into groups of control, ischemia/reperfusion and bFGF-treated, randomly. Apoptotic cells were detected using the TdT-dUTP terminal nick-end labeling at 1 hour, 6 hours, 12 hours, 24 hours, and 72 hours after reperfusion. The expression of caspase-3 at specified times was determined by Streptavidin Peroxidase immunohistochemistry. Atomic absorption spectrum method was used to evaluate the intracellular calcium changes of retinal tissues., Results: In ischemia group, apoptotic cells began to appear at 6th hour after reperfusion and increased progressively with time. The number of apoptotic cells reached the peak 24 hour after reperfusion, and no apoptotic cells could be found at 72 hours. Changes in caspase-3 expression followed a similar trend. The intracellular calcium level of rat retina began to increase at 1 hour after reperfusion, and continued to increase with the reperfusion time. At 24 hours after reperfusion the intracellular calcium level reached the peak, and decline thereafter up to 72 hours. The patterns of change of the three markers of treatment group were similar to the above. However, the magnitude of changes was relatively lower. A statistically significant difference (P < 0.05) between the ischemia group and treatment group at 6th, 12th and 24th after reperfusion was observed., Conclusion: Apoptosis may play a vital role in ischemia-reperfusion injury of the retina. bFGF may have a therapeutical effect on ischemia-reperfusion injury by inhibiting the increase of retinal intracellular calcium stores and caspase-3 protein expression.

Objective: To study the distribution and expression of type IV collagen (Col IV), laminin (LN) and fibronectin (FN) in the basement membrane of malignant eyelid tumors so as to explore the relationships between the basement membrane and the degree of differentiation, metastasis and prognosis of malignant tumors., Methods: Eighty-five paraffin-embedded specimens of malignant eyelid tumors including basal cell carcinoma (BCC), squamous cell carcinoma (SCC) and sebaceous cell carcinoma were studied to identify the Col IV, LN and FN with immunohistochemical SABC techniques., Results: A continuous intact basement membrane (BM) could be identified in all of the normal tissues and benign tumor specimens. In the malignant tumor specimens, the BM was defective or absent. The positive expression rate of Col IV in BCC, SCC and sebaceous cell carcinoma was 65% (24/40), 56% (16/25) and 40% (8/20). The positive expression rate of LN was 58% (23/40), 40% (10/25) and 35.% (7/20), respectively. The positive expression rate of FN was 33% (13/40), 36% (9/25) and 30% (6/20), respectively. The positive expression rate of these extracellular matrices was correlated with the degree of differentiation of the tumor. The difference of positive rate between highly-differentiated and poorly-differentiated tumors was significant (P < 0.05). In a short term follow-up (6-18 months), a statistically significant correlation (P < 0.05) could be observed between tumor recurrence and BM integrity. A higher recurrence rate and early recurrence were found in patients with interrupted BM., Conclusion: The absence of expression of Col IV, LN and FN was related to the invasive capacity and recurrence of the tumor. This result can be used as an index for clinical judgment of prognosis of the eyelid tumors.