Objective To investigate effects of different doses of recombinant human bone morphogenetic protein-2 (rhBMP-2) on vascular endothelial growth factor (VEGF) expression in fetal mouse osteoblasts.Methods Calvaria osteoblasts of fetal mice of 19 days were cultured.The effects of rhBMP-2 at different doses and different action times on VEGF expression patterns in fetal mouse osteoblasts were observed with reverse transcrip- tion-polymerase chain reaction (RT-PCR) and Western blot staining.Results In the present study,RT-PCR detected a steady expression of VEGF mRNA in the control fetal mouse osteoblasts.The levels of VEGF mRNA increased in an apparent biphasic manner with a maximum stimulation (about 2-fold above the control,P＜0.05 ) in both VEGF mRNA species observed at 300 ng/mL of rbBMP-2.After 48 h of rhBMP-2 treatment,the VEGF mRNA levels approached those in the control.The VEGF mRNA levels appeared to be biphasic in rhBMP-2-treated cultures,showing peak induction at 3 and 24 h and remaining elevated at 48 b.Compared with the individual control value at each time point,an apparent maximum increase (about 2.5-fold above the control,P＜0.05) occurred at 6 h.The second peak induction,about 2-fold above that in the control,occurred at about 36 h.Conclusions The expression of VEGF mRNA is steady in the control fetal mouse osteoblasts.RhBMP-2 can promote the expression of VEGF in dose-dependent and time-dependent manners.

Objective To explore the role of monitoring the cellular immune function in preventing and treating the fungal infection in the recipients of liver transplantation. Methods 679 cadaveric liver transplantations (from Jan. 2004 to Jan. 2010) were retrospectively studied. All the cases were divided into 3 groups according to different treatments and preventing regimens. The patients in groups A, B, C were treated based on the clinical experiences (394 cases), T lymphocyte subsets counting (151 cases), and combination of ATP values of CD4+ T cell and T lymphocyte subsets counting (134 cases), respectively. The infection, mortality and acute rejection rate were analyzed. The relationship between fungal infection and cellular immune function was investigated.Results The fungal infection rate in groups A, B, and C was 28. 9 %, 21.2 %, and 19. 4 % (P<0. 05), the morbidity rate was 16. 7 %, 12. 5%, and 3. 8% (P>0. 05), and the acute rejection rate was 28. 4 %, 17. 2 %, and 13. 4 % (P<0. 01), respectively. The CD4+ T lymphocyte counting in all cases of fungal infection was (147±43)×106/L. The morbidity could reach 50. 0 % when the CD4+ T lymphocyte counting < 100 ×106/L, while it was 2. 4 % when the counting was between (100-200) ×106/L (P<0. 01). The CD4+ T lymphocyte counting had no linear relation with the ATP value.The ATP value in fungal infection cases was (117 ± 61)μg/L. Conclusion The cellular immune function test could be quantitatively evaluated according to the T lymphocyte subsets and ATP value of CD4+ T lymphocyte. And individualized immunosuppressive therapy could be adjusted accordingly.Therefore, cellular immune function could be instructive in preventing and treating the fungal infection after liver transplantation.

Objective To explore the potential of breast cancer stem cells in differentiating into vascular endothelial cells and participating in the angiogenesis of breast tumor.Methods The expressions of mutant P53, CD31 ,VEGF and Her-2 amplification in tumor blood vessels were detected and the single cell suspension of breast cancer was prepared.CD44+/CD24-/low cells were selected and cultured in culture system.The expressions of CD31 and CD105 on endothelial cell surface were detected by EGM-2 endothelial cell culture medium.The ability of uptake of acetylated low-density lipoprotein was measured.The endothelial cells were cultured in vitro to observe their vascular structure.Results The expressions of CD31,VEGF and mutant P53 were found in paraffin sections of breast cancer tissue samples, and they were arranged along the vessel lumen or blood vessel sphere. Immunofluorescence showed that CD31 and DAPI signals were expressed intravascularly in breast cancer tissues. Four samples of Her-2 positive amplification and 9 cases of non-amplification were detected in the samples.The successful separation of breast cancer stem cells was carried out by immunomagnetic sorting.The percentage of CD44+ cells in the pre-sorting cells was (7.5±2.6)% and that of CD44+ cells was about (94.3±4.7)% after magnetic sorting (P<0.05).The ratio of CD24+ cells was (48.2±9.4)% before sorting,and that of CD24+ cells was (4.3±1.6)% after immunomagnetic bead sorting.The proportion of CD105+ cells in mammary gland cells was (4.5±0.9)% and the proportion of CD31+ cells was (6.2±1.3)%.After cultured for three generations,the percentages of CD105+ cells and CD31+ cells were (79.6±9.3)% and (84.1±10.7)%,respectively (P<0.05). DiL-Ac-LDL could be phagocytized by endothelial cells cultured in endothelial cell culture system.Vessel-like structure was found in the endothelial cell group while the control group did not have the tendency of vascular changes.Conclusion Breast cancer stem cell-derived endothelial cells differentiate into endothelial cells in vitro and have the ability of vascular formation,suggesting that breast cancer stem cells may be involved in the formation of tumor blood vessels.

AIM: To explore the histopathologic changes of Reis-Bücklers corneal dystrophy ( RBCD) . · METHODS: Cornea buttons were obtained from patients in 1 pedigree who underwent lamella keratoplasty. Sections with HE and special staining which included symplectic blue staining and Masson staining and Congo red staining were observed under light microscope. Two normal cornea specimens ( donated corneas from eye bank) were used as control. ·RESULTS:In those patients, the bowman's membrane disappeared. The main lesion was in bowman's membrane through HE staining, positive through PAS Congo red staining and Masson staining, negative through symplectic blue staining. ·CONCLUSION: The Reis-Bücklers corneal dystrophy is characterized as geographic map-like lesion of bowman's membrane, and the abnormal extracellular deposit is amyloid protein fiber.

AIM:To evaluate the short-term efficacy of posterior chamber phakic intraocular lens implantation for moderate to high myopia.METHODS:Fifty-three eyes of 27 patients with moderate to high myopia (-3.25 to-21.5D) were received posterior chamber phakic intraocular lens implantation with 2.8mm clear corneal incision.Changes of visual acuity, intraocular pressure(IOP), counts of endothelial cells and surgically induced astigmatism at 1mo after surgery were observed.RESULTS:The preoperative best corrected visual acuity (BCVA LogMAR) were 0.05±0.18, the postoperative BCVA at 1d were 0.02±0.14, the postoperative BCVA at 1wk were-0.05±0.11, the postoperative BCVA at 1mo were-0.07±0.10.The postoperative BCVA were statistically significant differences in different time(P<0.05).The preoperative endothelial cell count was 2932.5±270.8/mm2, the postoperative endothelial cell count at 1d was 2917.1±299.0/mm2;the postoperative endothelial cell count at 1wk was 2902.5±288.6/mm2;the postoperative endothelial cell count at 1mo was 2855.3±284.2/mm2.The postoperative endothelial cell count was not statistically significant differences in different time.The preoperative IOP was 15.8±2.5mmHg;the postoperative IOP at 1d was 15.3±2.9mmHg;the postoperative IOP at 1wk was 17.4±5.8mmHg;the postoperative IOP at 1mo were 15.8±2.6mmHg.The preoperative IOP was significantly different compared with postoperative IOP at 1d, 1wk and 1mo.The postoperative IOP at 1d and 1wk were statistically significant differences (P<0.05).The surgically induced astigmatism at 1d was 0.74±0.39D;the surgically induced astigmatism at 1wk was 0.57±0.28D;the surgically induced astigmatism at 1mo was 0.44±0.21D.The postoperative IOP were statistically significant differences in different time (P<0.05).The preoperative anterior chamber depth was 3.26±0.25mm;the postoperative central vault at 1d were 654.5±345.2μm;the postoperative central vault at 1mo were 683.0±211.7μm.The postoperative central vault were significantly different between that at 1d and 1mo after operations.Operations went without serious complications.CONCLUSION:Posterior chamber phakic intraocular lens implantation for moderate to high myopia were safe, effective and predictability.But long-term effect remains needed to be further follow-up.

Adult ; Female ; Humans ; Magnetic Resonance Imaging ; Neuroma, Acoustic ; pathology ; surgery

Aim To investigate the effects of irbesartan on Wnt/β-catenin signaling pathway in tubular epithelial-mesenchymal transition(EMT)in HKCs induced by high-glucose.Methods Human kidney proximal tubular epithelial cell line(HKCs)cultured in vitro was divided into four groups:normal-glucose group,mannitol control group,high-glucose group and high-glucose plus irbesartan group.Immunocytochemistry staining was used to observe the expression of β-catenin;the protein expression of Wnt4,β-catenin,E-cadherin and α-SMA was assessed by Western blot;Wnt4 and β-catenin mRNA were detected by reverse transcription-polymerase chain reaction(RT-PCR).Results Compared with normal-glucose and mannitol control group,both the protein and the mRNA of Wnt4 were up-regulated in HKCs stimulated by high-glucose.α-SMA expression significantly increased but E-cadherin decreased in HG group.The cytoplastic and nuclear fraction of β-catenin enhanced with highglucose stimulation.But no difference of the total protein and mRNA of β-catenin was observed between highglucose-treatment and control groups.Highglucose induced Wnt4 and β-catenin expression in a time-dependent manner,both peaking at 24 h.Irbesartan reduced the promotional effect of HG on Wnt4 and α-SMA expression,and nuclear translocation of β-catenin.HG-mediated inhibition of E-cadherin was also restored by irbesartan.Conclusion These data supported a functional role for Wnt/β-catenin signaling pathway during epithelial-mesenchymal transition of HKCs induced by high glucose and suggested that irbesartan might reverse tubular EMT by regulating activity of Wnt/β-catenin pathway.

Objective To explore the monitoring and the individualized adjustment of cellular immunology function in the recipients infected with pan-drug resistant Acinetobacter baumannii(PDR-Ab)after liver transplantation.Methods We retrospectively summarized the infection and the prognosis of PDR-Ab in 299 cases of liver transplantation performed from Jan.2008 to May 2010.The absolute number of T lymphocytes and ATP level within CD4+ T cells were monitored,and T cell immunology function(TCIFS)was scored.According to different immunology adjusting proposals,14 cases of PDR-Ab infection were divided into 2 groups:(1)traditional group,routine anti-infective therapy;(2)individualized group.Individualized immunology adjustment was made according to the score of TCIFS besides routine therapy.Results There was no significant difference in age,MELD and Child-pugh score between two groups.The peri-operative bleeding volume in individualized group was more than that in traditional group(P<0.01).There was no significant difference in TCIFS score between two groups at 1st week after transplantation and the onset of the PDR-Ab infection.However,the score in individualized group was apparently higher than that in traditional group when anti-infection therapy ended(P<0.05).The difference in the recovery rate between two groups was significant(P<0.05).No rejection happened in two groups.Conclusion It is an effective way to decrease the mortality of PDR-Ab infection after liver transplantation that the individualized adjustment of immunosuppression protocols is guided by grading quantitatively the cellular immunology function according to the absolute number of T lymphocytes and ATP level within CD4+ T cells.

To analyze dryeye disease after the femtosecond laser and mechanical microkeratome for laser in situ keratomileusis ( LASlK ) by Oculus dry eye analyzer examination.METHODS: The people who had femtosecond LASlK ( FS-LASlK ) and mechanical microkeratome LASlK in Weifang Eye Hospital from January 2014 to April 2014 were observed. Sixty patients ( 120 eyes ) who had corneal refractive surgery were collected. The average age of patients was 27. 46 ± 6. 65 years old. The LASlK group had 28 patients ( 56 eyes ) and the FS-LASlK group had 32 patients ( 64 eyes ) . The tears river altitude, first tear break up time ( BUT) , and average BUT at preoperative, 1wk and 1mo after operation were observed by Oculus dry eye analyzer.RESULTS:By Oculus dry eye analyzer examination, the preoperative tears river altitude of patients was 0. 25 ± 0. 10mm. First BUT was 6. 70 ± 3. 86s. Average BUT time was 10. 14 ± 4. 51s. At 1wk after operation, the tears river altitude of patients was 0. 19±0. 04mm. The first BUT time was 4.87±1.71s. The average BUT time was 7.77±2.30s. There was significant difference between preoperative and 1wk after operation (P<0. 05). At 1mo after operation in two groups, the tears river altitude of patients was 0. 19 ± 0. 18mm . The average BUT 8. 72 ± 2. 54s. The BUT was 5. 02 ± 2. 30s. There was significant difference between preoperative and 1mo after operation (P<0. 05). The two groups had significant difference in average tears river altitude and average BUT between 1mo and 1wk after operation. There was no significant difference in the first BUT between 1mo and 1wk after operation. There was no significant difference between LASlK and FS - LASlK groups at any time of the test (P<0. 05).CONCLUSlON: LASlK and FS - LASlK patients have different degree of dry eye symptom after operation. Dry eye symptoms mainly display in the decrease of tear production and the stability of tear film. lt will be better after giving artificial tears. Oculus dry eye analyzer is a non-invasive examination equipment. lt can reflect the amount of tears and the stability of tear film. lt is usefull in rapid diagnosis of dry eye and judgment of the degree of dry eye symptom after LASlK operation.

Seventy-two endophytic fungi isolated from Cephalotaxus hainanensis Li were screened for antitumor and antimicrobial activities. The results showed that 9 strains presented cytotoxic activities to at least one of the tested tumor cells, five strains exhibited strong antimicrobial activity against Staphylococcus aureus, one strain had inhibitory effect on Phytophthora parasitica. It was evidently that the endophytic fungi of C. hainanensis are potential resources to find valuable bioactive components.