AIM: To establish the co-culture mode of tissue-engineered tracheal seeding cells, and compare with the routine culture, so as to provide the fundament of tissue-engineered trachea construction by cell compound materials. METHODS: The experiment was completed at the laboratory, Department of Respiratory Diseases, Tangdu Hospital, the Fourth Military Medical University of Chinese PLA from May 2006 to May 2007.①Three healthy New Zealand male rabbits, weighed (250.00?0.75) g, were used in this study.②Co-culture mode: The tracheal epithelial cells and fibroblasts were isolated and then co-cultured for 7-10 days. The two kinds of cells were distinguished according to their different tolerances to trypsin. Subsequently, A hole received 0.5 g/L trypsinization, cells were suspended in DMEM medium containing 0.05 volume fraction of CO2 and then transplanted into B hole (fibroblasts); again, A hole was added with 2.5 g/L trypsin for digestion, afterwards cells were suspended with K-FSM medium and transplanted into C hole (tracheal epithelial cells). Routine culture: after isolation, tracheal epithelial cells were cultured and fibroblasts were purified, respectively.③The co-cultured cells and the traditionally cultured cells were compared via cell growth curve and cell proliferation detected by 3-(4,5-dimethylthiazo-2-yl)-2,5-diphenyl-tetrazolium bromide (MTT) assay. RESULTS: ①Cell growth: Co-cultured cells grew well, those in A hole increased intercellular apace after trypsinization, while cells in B hole were shaped as fusiform, without the manifestation of typical road stone-like cells, and cells in C hole were apposite, showing lamellar road stone-like morphology.②Cell growth curve: The growth curve of co-cultured cells was identical with that of cells by routine culture method.③Cell proliferation: MTT detection revealed no significant difference in the proliferation of co-cultured cells and cells cultured by traditional method (P

Objective To study the effects of Pioglitazone on TLR4 expression and peritoneal macrophage foam cells formation induced by ox - LDL.Methods Foam cells were cultured equally divied into four groups in random:the normal control,pioglizaone group,ox-LDL group and ox-LDL +pioglizaone group.The SD rats peritoneal macrophages were pretreated with ox - LDL(50 mg/mL)and Pioglitazone(20 mmo1 /L)respectively for 24 h.Then they were stained by oil red O to determine foam cells formation.The effects of Pioglitazone on foam cells TLR4 ex-pressions were detected by immunohistochemistry.Results The SD rats peritoneal macrophages of control group and Pioglitazone group are not almost stained by oil red O,but the macrophages of ox-LDL group become dark red after stained by oil red O,and the cells color of ox-LDL + Pioglitazone group is thinner than that of ox-LDL group.The expressions of TLR4 in ox-LDL group are higher than those in control and Pioglitazone group significantly(24 h,t =3.015,t =4.007;48 h,t =3.100,t =4.008,P <0.01).while expressions in ox-LDL +Pioglitazone group are lower than ox-LDL group respectively(24 h,t =4.075;48 h,t =4.150,P <0.05).Conclusion Pioglitazone might inhibit the formation of foam cells by suppressing the expressions of TLR4 in macrophages.

Objective: Through summarizing the main problems and challenges faced in the medical health system reform in China , suggestions are given: the government needs to erhance the health investment and play the role of market mechanism , make the use of electionical information technique, further change the community system of implementing medicine community system, implement the medical service supervision system.

Objective To evaluate clinical effects of the transurethral ureteroscopic lithotripsy(URL) and minimally invasive percutaneous nephrolithotomy(MPCNL) in the management of upper ureteral stones.Methods Clinical data of 258 patients diagnosed as having upper ureteral calculi from January 2001 to December 2004 in this hospital were retrospectively analyzed.The patients were given either URL(225 patients) or MPCNL(33 patients).Results Of the URL,the stone-free rate on one session was 73.8%(166/225) and the failure rate was 26.2%(59/225).The causes for the failure included stone movement to the renal pelvic in 42 patients(18.7%,42/225),unsuccessful manipulation in 14 patients(6.2%,14/225),and conversions to open surgery because of ureteral perforation in 1 patient and ureteral rupture in 2 patients(1.3%,3/225).Of the MPCNL,the stone-free rate on one session was 100%(33/33).Conclusions The application of URL can be interfered with ureteral stricture and twist that are secondary to incarcerated ureteral stones.High stone-free rate,low incidence of complications,and satisfactory reliability can be expected using MPCNL,especially in patients with impacted ureteral calculi accompanied with secondary affection at the same side.

Objective To observe the influence of early postoperative feeding in the healing of intestinal anastomosis in rabbits,and to clarify preliminarily the relationships between early postoperative feeding after gastrointestinal surgery and gastrointestinal anastomotic fistula formation and healing time in rabbits.Methods 48 rabbits were randomly divided into experimental group and control group, then they were treated with gastrointestinal anastomosis.The rabbits in experimental group were fed with liquid diet 24 h after operation,and the rabbits in control group were fed nothing after operation and supplied by total parenteral nutrition.Two rabbits of each group were selected for exploratory laparotomy on the 3rd,5th,7th,10th and 15th day after operation,and the healing rate of anastomosis,the anastomotic bursting pressure,the anastomotic breaking strength,and the hychoxyproline level of anastomosis were observed.Results The healing rate of anastomosis in control group was 91.6%(22/24), and the healing rate of anastomosis in experimental group was 95.8%(23/24),there was no significant difference between two groups(P>0.05).The anastomotic bursting pressures of the rabbits in two groups were decreased remarkably at the 72nd hour after operation,which was the lowest point,and they were increased remarkably on the 5th day after operation,but the anastomic bursting pressure in experimental group was a little lower than that in control group,and it reached the peak on the 7th day after operation in control group. On the 10th day after operation,the anastomic bursting pressure in control group was a little lower than that on the 7th day after operation,but the anastomic bursting pressure in experimental group reached the peak.There were no significant differences of anastomic bursting pressure at different time points between two groups(P>0.05).The anastomotic breaking strength had no significant difference between two groups at the 72nd hour after operation,both of them reached the lowest points,however the anastomtic breaking strengths in two groups were increased remarkably on the 10th day after operation,and reached the peaks.but there were no significant differences of anastomic breaking strength at different time points between two groups (P>0.05 ). The hychoxyproline level of anastomosis:in experimental group was a little lower than that in control group at the 72tnd hour after operation,and both of them reached the peaks on the 7th day after operation;but there were no significant differences of hychoxyproline levels of anastomosis at different time points between two groups(P>0.05).Conclusion Early postoperative feeding can not cause the increase of anasmotic healing time and the incidence rate of gastrointestinal anastomotic fistula.

AIM: To observe the effect and safety of applying mitomycin C ( MMC ) in laser - assisted subepithelial keratomileusis ( LASEK) for extreme high myopia.
●METHODS:LASEK with 0. 02% MMC was performed in 364 eyes of 182 patients with extreme high myopia and spherical equivalent was-9. 0 to-10. 25 D. All the patients were divided into four groups according to the applying time of MMC. The time in Group l was 10s, in GroupⅡwas 25s, in Group Ⅲ was 40s and 55s in Group lV. Uncorrected visual acuity ( UCVA ) , residual refraction, Haze, healing time of corneal epithelium, density and variant index of corneal endothelium was examined in these patients and follow-up 6mo.
●RESULTS:At 6mo after surgery, 76. 3%, 94. 0%, 92. 3%and 93. 8% of the patients in Group l, GroupⅡ, GroupⅢ and Group lV respectively achieved the UCVA better than 15/20. There was significant difference of UCVA between Group l and the other three groups (χ2=19. 610, P=0. 000). Proportion of the residual refraction between ± 0. 5D in Group l (78. 8%) was lower than other groups (95. 2% in Group Ⅱ, 93. 3% in Group Ⅲ, and 92. 7% in GroupⅣ) at 6mo and there was significant difference (χ2=16. 329, P=0. 001). Group l had more Haze statistically than the other three groups at 6mo postoperatively ( Hc=50. 110, P=0. 000). The healing time of cornea epithelium seem to be no statistically difference between each group at 6mo (χ2=11. 611, P>0. 05). MMC had no influence on the density of corneal endothelium in each group postoperatively (P>0. 05), there were 3071. 3±284. 4 cells/mm2 in Group l, 3 105. 6 ± 337. 8 cells/mm2 in Group Ⅱ, 2 986. 3 ± 304. 1cells/mm2 in Group Ⅲ and 3 088. 7 ± 372. 5 cells/mm2 in Group lV respectively. The variant index of corneal endothelium calculated in each group at 6mo after surgery was 24. 72 ± 6. 52, 22. 93 ± 6. 74, 24. 38 ± 6. 63 and 23. 14±7. 22 repectively, compare with that preoperatively there were no statistically differences (P>0. 05).
●CONCLUSION: For extreme high myopia LASEK with 0. 02% MMC is effective and safe. The MMC applying time of 25s in LASEK can effectively reduce Haze after surgery and decrease potential complications.

AlM:To make a analysis of visual quality of iris location guided femtosecond laser assisted in situ keratomi ( LASlK) and iris location guided mechanical sub-bowman keratomileusis ( SBK) for high myopia treatment.METHODS:Femtosecond LASlK ( study group ) was performed in 102 eyes of 51 patients with high myopia and 70 eyes of 35 patients were received mechanical SBK ( control group ) from January to October 2013. The spherical refraction of all the patients was from -6. 00 ~-9. 50D. Best corrected visual acuity ( BCVA ) of the patients was ≥1. 0. Uncorrected visual acuity ( UCVA), BCVA, thickness of cornea flap, contrast sensitivity function ( CSF ) and senior ocular aberration were examined in these patients and follow-up was 1a.RESULTS: At 1a after surgery 94. 1% UCVA in study group reached ≥1. 0 and there was 94. 3% in control group. There was no significant difference between two groups (P>0. 05). Residual refraction of study group was-0. 08±0. 10 D and control group was -0. 10±0. 07 D. There was no significant difference of residual refraction between two groups (P>0. 05). C12 , C8 of senior ocular aberration and RMSH in study group was less than control group, amplification:0.1642±0.0519 and 0.2229±0.0382 (t=8. 077, P<0. 05), while C7 was equal in two groups:0. 1583 ±0.1921 and 0. 1436±0. 1536 (t=0. 556, P>0. 05). C8 was 0. 0950±0. 069 and 0. 1858±0. 095 (t=7. 261, P<0. 05). RMSh was 0. 3711 ± 0. 0847 and 0. 5622 ± 0. 1033 ( t= 12. 801, P<0. 05). CSF was no significant difference between study group and control group (P>0. 05). CONCLUSlON: Femtosecond LASlK and mechanical SBK is effective for high myopia. Compared to mechanical SBK, femtosecond LASlK shows more advantages in the senior ocular aberration and visual quality. The cornea flap is more regular from central to peripheral area with femtosecond laser.

Objective To study the effect of sensory nerves on the expression of epidermal growth factor(EGF), basic fibroblast growth factor(bFGF) by fibroblasts in the granulation tissues in skin wound healing. Methods In vivo and in vitro studies were conducted. Fifty wistar rats were randomly divided into capsaicin group and control group. The rats in the capsaicin group were subcutaneously injected with neurotoxin dose of capsaicin(50 mg/kg for 3 days) to chemically destroy the sensory nerves. Full thickness skin incisive wound was made on the rat′s back on 8th day after last injection. The contents of substance P(SP), which was mainly released from afferent sensory nerve terminal, and EGF, bFGF in granulation tissue of skin lesion were determined using immunohistochemistry combined with computerized imaging analysis on 3rd, 6th, 9th, 12th days after the incision. In the control group no capsaicin was injected. In the in vitro study the fibroblasts from granulation tissue of rats were cultured with different doses of SP(10-9 ~ 10-5 mol/L), and the protein expression of EGF, bFGF by cultured fibroblasts was determined using Western blot. Results Immunoreactive SP was not only found around the small blood vessels and in the intercellular space, but also in the cytoplasm of fibroblasts in granulation tissue. SP content and the protein expression of EGF, bFGF in granulation tissue decreased in the capsaicin group, compared with those in the control group, which indicated that SP(10-7 mol/L) added into the culture medium for fibroblasts could upregulate the levels of EGF, bFGF expression. Conclusions Protein expression of EGF and bFGF by fibroblasts in granulation tissue in skin wound healing might be related with the SP released from sensory nerves.

Objective To evaluate the effect of surgical treatment in facial nerve paralysis.Methods Clinical data of 29 cases in facial nerve paralysis were retrospectively analyzed.All of the 29 cases of facial paralysis,18 cases is the suppurative otitis media,9 cases is the temporal bone fracture,2 cases is the neoplasms of the temporal bone.The 29 cases of facial nerve paralysis were surgical treatment.8 cases by vertical segment or horizontal segment of facial nerve decompression,19 cases by the stylomastoid foramen to the geniculate ganglion of facial nerve decompression,2 cases by the itratemporal course of facial nerve decompression.1 case was underwent end-to-end anastomosis,2 cases of the greater auricular or the sural nerve graft for repairing facial nerve defect.All data were analyzed with Rank sum test.Results Makes a follow-up visit for 6～18 months,the facial nerve function(House-Brackman grading system)before the technique Ⅱ 6.9%,Ⅲ17.2%,Ⅳ34.5%,Ⅴ 31.0%,Ⅵ 10.3%,after the technique,restores Ⅰ 6.9%,Ⅱ27.6%,Ⅲ27.6%,Ⅳ 24.1%,Ⅴ 13.8%,statistics analysis facial nerve function restoreS has the significance difference(P＜0.005).Conclusions The facial nerve decompression and the nerve graft are useful method to treat facial paralysis.Surgical treatment of facial paralysis is satisfied in the suppurative otitis media and the temporal bone fracture.

Objective To compare the influence of general anesthesia and spinal anesthesia on the mortality in rats with fat embolism.Methods Three hundred and seventy five rats were randomly assigned to receive general anesthesia （group GF,n=125）,spinal anesthesia （group SF,n=125）,or no anesthesia （group F,n=125）.Rats were injected with a half lethal dose of fat from caudal vein 20 min after anesthesia induction. The mortality was recorded at 2,8,12,and 24 h after fat injection,respectively.Results The mortality was lower in group GF than those in groups SF and F （P0.05）,while there was no statistical difference between group SF and group F.Conclusion The mortality in rats with fat embolism under general anesthesia was lower than that under spinal anesthesia.