Objective To investigate the efficacy,safety and localization of multilayer amniotic membrane transplantation(AMT) for corneal perforation associated with ulceration.Design A retrospective clinical case series.Participants Nine patients(9 eyes)with corneal perforation secondary to ulceration were enrolled into this study.These were little response to medicine,including bacterial keratitis(4 eyes ),fungal keratitis(2 eyes),fungal and bacterial mixed keratitis( 1 eye),virus keratitis( 1 eye),and Mooren's ulcer ( 1 eye).Size of perforation was 0.5～3.0 mm in diameter.Methods The AMT was applied to tamp the perforation,fill the ulcer and cover the surface of ulcer.After surgery,the medicine was continued to be used to treat the original corneal ulcer.The follow-up ranged from 6～20 months.In the suffering eyes,reformation of anterior chamber,healing of ulcer,complications and recurrence of ulceration were observed.Main Outcome Measures Of the postoperative eyes,reformation of anterior chamber,healing of ulcer,complications and recurrence of ulceration.Results The anterior chamber reformed at the first postoperative day in 9 eyes,and kept in normal depth in the follow-up time.At postoperative 2 months,the ulcer healed with sear and a smooth surface.Corneal thickness of the ulcer area recovered almost to normal.During follow up,no recurrence of ulceration or severe complications was noted.Conclusions Multilayer AMT is a safe and effective method for the management of small corneal perforation associated with ulceration,but the ulcer healed with scar.(Ophthalmol CHN,2008,17:101-103)

AIM: To investigate the effect of tetrandrine (Tet) on proliferation of human pterygium fibroblasts (HPF) in vitro and to search for a new method to prevent the recurrence after pterygium surgery.METHODS: With different concentrations (0 to 160μmol/L) of Tet acting on HPF cultured in vitro, the impact was observed at 24, 48, 72 hours respectively after Tet intervention. The MTT method was used to assay the biologic activities of Tet and inhibitive rate of cell growth. The expression of proliferating cell nuclear antigen (PCNA) in each group was detected by immunohistoche-mistry before and after Tet intervention.RESULTS:With different concentrations of 20, 40, 80 and 160μmol/L and acting for 24 to 72 hours, Tet could inhibit the proliferation of HPF in a dose- and time-dependent manner (P<0.05). After the intervention of Tet, the expression of PCNA protein declined. When the concentration of Tet was in the range of 20 to 160μmol/L, it was able to inhibit the expression of PCNA in a concentration-dependent manner (P<0.05).CONCLUSION:Tet could significantly inhibit the proliferation of pterygium fibroblasts, and the inhibitive action was in a dose- and time-dependent manner within a certain range of concentration. But in high concen-tration (>160μmol/L), Tet would have cytotoxity.

AIM: To investigate the efficacy, safety and localization of multilayer amniotic membrane transplantation (AMT) for the management of corneal perforation associated with ulceration.METHODS: Six eyes (6 patients) were treated with multilayer AMT for corneal perforation associated with ulceration. The perforation size was 0.5-2mm in diameter. The amniotic membrane (AM) was applied to tamp the perforation, fill the ulcer and cover the surface of ulcer. The follow-up ranged from 5-19 months.RESULTS: The anterior chamber reformed at the first postoperative day in all patients, and kept in normal depth in the follow- up time. The epithelialization of the AM grafts was observed in about 3 weeks after surgery. During the first postoperative month, the corneal thickness at the perforation site gradually increased, and the stromal inflammatory cell invasions surrounding the corneal ulcer were gradually subsided.Two months postoperatively, the ulcer healed with scar with part neovascutarization; the corneal thickness of the ulcer area was stable and recovered almost normal. The vision was not improved in all patients after operation. During-follow up,no severe complications or recurrence of ulceration was noted.CONCLUSION: Multilayer AMT is an effective and safe method for the management of small corneal perforation associated with ulceration, but the ulcer healed with scar.

AIM: To evaluate the efficacy and safety of annular bubble conjunctival excision combining with amniotic membrane transplantation(AMT) for vernal keratoconjunctivitis(VKC) with gelatinous swellings(papillae) at the limbus.METHODS: Twenty eyes(10 patients) with VKC, were characterized by gelatinous swellings(papillae) at limbus and were underwent annular bubble conjunctival excision associated with AMT. The follow up ranged 5-21months.RESULTS: After operation, the symptoms were subsided remarkably: itching,discharge and foreign body sensation were almost disappear; photophobia and tearing were obvious in the first week after surgery, but were gradually alleviated, and almost disappear after one month of postoperative.Yellow-brown or filthy red gelatinous protuberances at limbus were cleaned away by operation and did not recur during the follow up. Corneal superficial punctuate keratitis(11 eyes) and corneal ulcers(3 eyes) were healed during the first week after surgery and did not recur. Conjunctival congestion was gradually lessened after surgery, which could not be noted 1months after surgery in 19 eyes, and the conjunctiva around the limbus, where AMT was performed, was whiter than the normal. But conjunctival congestion in 1 eye lasted for about 5mon accompanying with a little of itch, and this eye was treated with eyedrops with improvement but recurrence. No severe complications had been seen during the follow up.CONCLUSION: Annular bubble conjunctival excision combining with amniotic membrane transplantation is an effective and safe method for the treatment of vernal keratoconjunctivitis with gelatinous swellings (papillae) at limbus. But the candidates must be chosen with discretion.

AIM: To investigate the efficacy and safety of annular conjunctival excision combined with amniotic membrane transplantation for the management of earlier Mooren's ulcer.METHODS: Thirty eyes (24 patients) with earlier Mooren's ulcer were treated with this method, followed-up for 5-30mo. The characteristic of these patients was localized periphery corneal ulcer: the ulcer involved 30-120° corneal limbus with depth of 1/3-1/2 cornealthickness and width of 2-5mm.RESULTS: After surgery, symptoms subsided remarkably and immediately in all patients,and almost disappeared after stitches were removed. Corneal melts were controlled. At postoperative 2-3mo, the corneal thickness of the ulcer area became almost stable, which in some patients recovered normal and in others were still thinner than normal, and the average increase was about 1/3 of the corneal thickness. Corneal transparence and vision were improved. During follow-up, no severe complications or recurrence were noted.CONCLUSION: Conjunctival peritomy combined with amniotic membrane transplantation is an effective and safe method for the treatment of earlier Mooren's ulcer.

Objective To preparegraphene quantum dots (GQDs) and construct a novel biosensor for determination of dopamine (DA) and uric acid (UA).Methods The GQDs was prepared by carbonization method from citric acid as carbon sources.Differential pulse voltammetry was used by the modified electrodes to detect uric acid and dopamine,and the detection performance was evaluate.Collected three experimenters morning urine on July 29,2016.The proposed sensor was used for biological samples detection.Results The size of GQDs were between 3 to 5 nm,which was observed by transmission electron microscopy (TEM).The proposed sensor had good linear correlation of 50～600 μmol/L UA (r2 =0.996 6) and 2～ 240 μmol/L DA (r2 =0.992 5).In uric acid in urine samples' detection (n=3),RSD value was less than 3％.The standard addition recovery rates of UA and DA were in 95.7％～101.4％ and 96％～102％ respectively.Conclusion The biosensor based on GQDs for determination DA and UA had been constructed successfully.

OBJECTIVE To investigate the positive rate of ?-lactamase,oprD2,aminoglycosides modifying enzyme and qacE△1 gene among clinical isolated strains of multi-resistant Pseudomonas aeruginosa in our hospital.METHODS P.aeruginosa was determined by VITEK,and MIC was determined by agar dilution method.TEM,IMP,VIM,oprD2,aac(3)-Ⅱ,aac(6′)-Ⅰ,aac(6″)-Ⅱ,ant(3″)-Ⅰ,ant(2″)Ⅰ and qacE△1 in strains were detected by polymerase chain reaction(PCR).RESULTS The positive rate of TEM,aac(3)Ⅱ,aac(6′)-Ⅱ,ant(3″)-Ⅰ and ant(2″)-Ⅰ in multi-resistant P.aeruginosa were 51.4%,48.6%,40.0%,54.3%,45.7% and 60.0%,respectively.No IMP and VIM genes were detected.CONCLUSIONS Positive rate of ?-lactamase, aminoglycosides modifying enzyme is high in 35 tested strains,and that should be paid more attention.

BACKGROUND: Hundreds of mouse embryonic stem cell lines are established presently. Most of these cell lines are collected from 129, C57BL/6J and BALB/C mice. Success rate of creating embryonic stem cell lines with Kunming mice is very low. OBJECTIVE: To explore the method of isolation and culture of murine embryonic stem cells of kunming species to increase the efficiency of establishment of embryonic stem cell lines from Kunming species mouse. DESIGN, TIME AND SETTING: The cell experiment was performed at the Chongqing Key Laboratory of Neurology from April 2006 to June 2007. MATERIALS: Blastocysts of 3.5 days and 4 days from Kunming species mouse were collected. METHODS: The blastocysts of 3.5 days and 4 days from Kunming species mouse were cultured on the fibroblast cell feeder layers for 4-5 days, and the cells from inner cell mass were isolated and subsequently cultured in vitro in 2.5 g/L trypase-0.4 g/L ethylenediamine tetraacetic acid (EDTA) and 1 g/L trypase-0.2 g/L EDTA at room temperature for 2-4 minutes. Embryonic stem cell colony was mechanically straggled. MAIN OUTCOME MEASURES: Colony growth was observed and determined by alkaline phosphatase staining, OCT-4 staining and karyotype analysis. RESULTS: Adherence rate, inner cell mass frequency and cloning efficiency of 4 day post coitum (dpc) embryo were higher than those of 3.5 dpc embryo (P

Objective To explore the effect of embryonic stem cells(ESCs) differentiating into cardiomyocytes(CM) by cardiomyocytes.Methods The blastocytes of 3.5 d from Kunming mouse,cultured on mouse embryonic fibroblaste cell feeder layers for 4～5 d,the mouse ESCs were isolated and subsequently cultured.Day 2～3 embryoid bodies(EBs) were derived from ESCs of passage 3 to 5 and then incubated with neonatal cardiomyocyte(CM) to induce into cardiomyocytes and the expression of cardiac specific cardiac troponin-T(TnT) and ?-actin were detected by immunofluoresence.Results The rhythmic beating embryoid bodies were observed on day 3 and reached 93% of rhythmic beating embryoid bodies on 12 day in group 2.All the beating cardiomyocytes derived from ESCs expressed cardiac-specific proteins for TnT and ?-actin,and the group induced by direct cell contact acquired the highest differentiating ratio as 56.5%,and was the highest differentiating ratio compared with other groups(P

Objective To explore the prognostic risk factors of low level malignant obstructive jaundice treated with transhapetic biliary drainage.Methods The clinical data of 142 patients with low level malignant obstructive jaundice received percutaneous transhapetic biliary drainage management from January 2010 to June 2013 were retrospectively analyzed.The study parameters included gender,age,tumor type,preoperative obstructive time,preoperative infection,drainage method,Child-Pugh grade,serum total bilirubin (TBIL),albumin (ALB),serum creatinine (SCr),the postoperative declining degree of bilirubin and postoperative antineoplastic therapy.The prognostic risk factors were evaluated.Results Single variable analysis showed that preoperative infection (P =0.006),Child-Pugh grade (P =0.004),SCr (P =0.043),the postoperative declining degree of TBIL (P =0.001) and postoperative antineoplastic therapy (P =0.015) were the related factors for survival time.The further Logistic regression analysis showed that preoperative infection (OR =3.729,95％ CI 1.332-6.363,P =0.040),Child-Pugh grade ≥ 10 scores (OR =0.513,95％ CI 0.375-1.276,P =0.018) and postoperative antineoplastic therapy (OR =0.668,95％ CI 0.210-2.026,P =0.038) were the related factors for survival time.Conclusion In treating of low level malignant obstructive jaundice with transhapetic biliary drainage,the preoperative infection,Child-Pugh grade and postoperative antineoplastic therapy may be the important related factors that affect the patient's survival time,to evaluate the prognosis of these patients has important reference meaning.