Objective To investigate the possibility to obtain the cranial subtraction computed tomography perfusion imaging (sCTP) from the CTP examination, and to explore preliminary application value of sCTP. Methods Cerebral sCTP obtained from source imaging data of 11 patients (10 with stroke and 1 with brain tumor) were analyzed with subtraction software and perfusion 3 software at GE AW4.2 workstation. The parametric maps and region of interest (ROI) value of blood flow (BF), blood volume (BV), mean transit time (MTT) and/or permeability surface (PS) were generated from the CTP and sCTP imaging data respectively. Results ①The technical success rate of cranial sCTP created from CTP sources imaging data was 100%, and the post-processing time of sCTP was about from 1.0 to 2.5 h. The mappings and values of BF, BV, MTT and PS of ROI were all successfully obtained from sCTP and CTP imaging data. ②The cerebral mappings of BF, BV, MTT and PS obtained from sCTP were similar to those from CTP, which all depicted accurately the lesions and their microcirculation hemodynamics changes. ③Just like those from CTP, the values of BF, BV, MTT and PS of nidus obtained from sCTP were significantly different from those of the near and the contralateral corresponding cerebral tissues (P<0.025). Conclusion sCTP can be generated successfully with subtraction and perfusion imaging software. It is an alternative solution for functional diagnosis of stroke and brain neoplasms.

Objective To evaluate the differences of the thinnest-point corneal thickness (TCT) decrease after three different corneal crosslinking (CXL) protocols for progressive keratoconus.Methyds Retrospective clinical case study.From August 2010 to November 2015,consecutive 85 patients (110 eyes) with progressive keratoconus were enrolled and treated with CXL in Department of Opthalmology,Navy General Hospital.21 patients of 25 eyes underwent standard epithelium-off corneal crosslinking (S-CXL),14 patients of 22 eyes underwent 1 g · L-1 riboflavin-sodium lactate Ringer's solution iontophoresis-assisted CXL (I-CXLa),and 50 patients of 63 eyes underwent 0.1％ riboflavin-distilled water solution I-CXLb.Preoperative and postoperative TCT were measured by ALLEGRO oculyzer.The differences of TCT decrease after treatment were compared among the three CXL protocols.Results The differences of TCT from baseline after 3 months,6 months and 12 months in the S-CXL group were (-14.93 ±27.16) μm,(-31.94 ±22.89) μm,(-27.71 ±26.01) μm,respectively,the I-CXLa group were (-20.14 ± 19.09) μm,(-10.10 ± 24.28) μm,(-7.11 ± 22.26)μm,respectively,the I-CXLb group were (-28.08 ± 26.14) μm,(-21.08 ± 25.62) μm,(-15.91 ± 19.19)μm,respectively.Three months after treatment,the differences of TCT decrease in the three groups was not statistically significant (P =0.188);Six and 12 months after treatment,the differences between S-CXL and I-CXLa were statistically significant (all P ＜0.05),but the differences between S-CXL and I-CXLb,between I-CXLb and I-CXLa showed no significant difference (all P ＞ 0.05).Conclusion Six and 12 months after treatment,TCT decrease is related to the CXL protocol.TCT decrease degree may reflect the intensity of crossinking.TCT decrease in I-CXLb is smaller than that in S-CXL,but there is no statistical difference.

Objective To evaluate the sensitivity of combination of itraconazole and amphotericin B to20clinical isolates of Cryptococcus neoformans and their in vitro interactions.Methods The sensitivity of combination of itraconazole and amphotericin B and their in vitro interactions were determined with20clinical isolates of C.neoformans by a checkerboard titration broth microdilution-based method in accord with the recommendations of the National Committee for Clinical Laboratory Standards(NCCLS),USA.Results When both drugs were given in combination,there was significant reduction of geometric means of MICs for itraconazole(from0.2730to0.1195?g/mL)and for amphotericin B(from0.6830to0.2102?g/mL).Synergistic effects were found in35%of isolates,additive effects in55%of isolates,and indifferent effects in10%of isolates.Antagonistic effects were not observed.The colony formation unit(cfu)per millilitre was significantly decreased in an isolate which was treated with different concentrations of the combination of both drugs,in comparison to that with the corresponding concentrations of individual drug.Conclusion The combination of itraconazole and amphotericin B is significantly more active against C.neoformans in vitro than individual drug alone.

Scanning electron microscopic observations were made on the morphological structures of the two copulatory spicules of the male Necator americanus and Ancylostoma duodenale. In both species,one of the two copulatory spicules was in the shape of a concave groove and the other, in the shape of an oblate tube. Owing to the difference in the concavity of the groove shaped copulatory spicule between the two species as shown by the cross sections, Necator americanus usually appear to have only one copulatory spicule whereas Ancylostoma duodenale usually show two separate copulatory spicules in appearance (Figs 1 -10).

Objective To investigate the technical protocols and feasibility of subtraction computed tomography perfusion imaging (sCTP) on observation of ischemic necrosis of femoral head (ANFH) on dog. Methods Sixteen laboratory canines underwent CT perfusion imaging (CTP) of femoral head before and after selective femoral circumflex artery embolization, and ANFH were observed. Then new sequence imaging data, created by sources imaging of CT perfusion scan using subtraction software, were analyzed at workstation (AW 4.2) with CT perfusion 3 analysis program, and data of sCTP were obtained. The parametric maps and indexes of capillary-level hemodynamics including blood volume (BV), blood flow (BF) and mean transit time (MTT) of CTP and sCTP were compared. Results ①The technical success rate of sCTP post-processing created from CTP sources imaging data was 100%. The values and mappings of BF, BV and MTT of region of interest (ROI) were all obtained from subtraction sequence images data. The post-processing time of sCTP was about 1-5 h. ② sCTP depicted ANFH well, though the values and mappings of BF, BV and MTT were different from those obtained with conventional bone CTP. Conclusion sCTP can be generated with subtraction and perfusion analysis program and techniques. It can be applied to bone CT perfusion imaging and early quantitative diagnosis of necrosis.

Background The incidence of dry eye is gradually increased,and researches showed that inflammation participated in the pathogenesis and development of dry eye.The current therapy for dry eye can only relieve symptom but not achieve final cure.Stem cell therapy has been used in the treatment of limbal stem cell deficiency.However,whether it is feasible for the stem cell treating dry eye is still unclear.Objective This study attempted to investigate a new approach to treat dry eye syndrom by using human umbilical cord mesenchymal stem cells (HUCMSCs).Methods The use and care of experimental animals complied with the Tsinghua University School of Medicine Laboratory Animal Care Details.HUCMSCs were cultured and cell suspension was prepared with the cell density of 5×105/ml.Twenty 24-week-old male NOD/Ltj mice were randomized into 4 groups.0.1 ml PBSHUCMSCs suspension was injected via tail vein or lacrimal respectively in the caudal vein injection group and lacrimal injection group,and 10 μ1 PBS-HUCMSCs suspension was topically administed in the eye drops group.The NOD/Ltj mice without any treatment served as the model group.Five male ICR mice were used as the normal control group.Tear secretion was quantitatively detected with phenol red cotton thread in 1,2,3 weeks after injection,and corneal epithelial defect was scored by fluorescein staining.The serum levels of interleukin (IL)-6,IL-17a,tumor necrosis factor-α (TNF-αt) and interferon-γ (IFN-γ) were assayed by ELISA.The relative expression levels of p65,Stat3,Stat5 and extracellular signal-regulated kinases (Erk)-1 in lacrimal gland were detected by Western blot.Results The tear secretion amount was significantly different among the model group,caudal vein injection group,lacrimal injection group and eye drops group in various time points (1 week:F =3.700,P =0.040;2 weeks:F =5.150,P =0.008;3 weeks:F=10.130,P＜0.001).The tear secretion amount was increased in the caudal vein injection group and lacrimal injection group compared with the model group in different time points (all at P＜0.05),and no significant difference was seen in tear secretion amount between eye drops group and model group among various time points (all at P＞0.05).The fluorescein staining score was 3.00±0.63,9.40±1.62,5.20±1.17,4.20±1.17 and 7.20±0.98 in the ICR mouse control group,model group,caudal vein injection group,lacrimal injection group and eye drops group 1 week after injection respectively,and the scores were significantly lower in the caudal vein injection group,lacrimal injection group and eye drops group than those in the model group (P =0.001,0.000,0.033).The serum levels of IL-6,IL-17a and TNF-α in the caudal vein injection group were evidently lower than those in the model group (t =4.70,3.46,11.0,all at P＜0.01),but no significant difference was displayed in the serum IFN-γ level among the five groups (F=1.740,P=0.170).The expressions of STAT5 were significantly decreased in the mice treated with tail vein injection and lacrimal injection compared with mice without treatment (both at P＜0.05).Conclusions Administration of HUCMSCs via intravenous and lacrimal injection can alleviate the inflammatory response during progression of dry eye syndrome by down-regulating the serum level and expression of inflammation-related factors in NOD/Ltj mice.The topical administration of HUCMSCs eye drops can attenuate the symptom of dry eyes by lubricating the cornea and suppling nutrition.

ObjectiveTo summarize the results of stereotactic radiation therapy (SRT) with or without whole-brain radiotherapy (WBRT) in the treatment of multiple brain metastasis.MethodsFrom May 1995 to April 2010,totally 98 newly diagnosed multiple (2 - 13 lesions) brain metastases patients were treated in our centre.Forty-four patients were treated with SRT alone and 54 with SRT + WBRT.Dose fractionation schemes were 15 -26 Gy in 1 fraction or 24.0 -52.5 Gy in 2 - 15 fractions with 3.5 - 12.0 Gy per fraction,depending on the tumor volume,location,and history of prior irradiation.Kaplan-Meier and Cox proportional hazards regression analyses were used for survival analysis.The median age of the whole group was 55 years.The survival time was calculated from the date of radiation treatment to the day of death by any cause.ResultsThe median follow-up time for the whole group was 12 months,and the follow-up rate was 100％.The median overall survival time was 13.5 months for the whole group,there was no difference between SRT alone group and SRT + WBRT group ( 13.0 months vs.13.5 months,χ2 =0.31,P =0.578 ).The Karnofsky Performance Score ( KPS) at the time of treatment ( χ2 =6.25,P =0.012 ),the interval between the diagnosis of the primary tumor and brain metastases ( χ2 =7.34,P =0.025 ) and the status of extracranial metastases ( χ2 =4.20,P =0.040) were independent prognosis factors for survival in multivariate analyses.ConclusionsStereotactic radiation therapy is an effective and alternative treatment choice for multiple brain metastases.

Objective To evaluate the role of nodal gene modulating malignancy of a hepatocellular carcinoma cell lines SMMC7721. Methods To silence the expression of nodal gene in human hepatocellular carcinoma cells by RNA interference ( RNAi),and to observe the effect on cells biological behaviour and vasculogenic mimicry.4 expression vectors of nodal gene targeting small interference RNA were constructed and transfected into SMMC-7721 cells.Real-time quantitive PCR and Western blot were used to examine nodal gene expression. The effects of nodal gene RNA interference on proliferation,apoptosis,infestation,migration and vasculogenic mimicry of SMMC-7721 were studed. Results The expression of nodal gene was suppressed in SMMC-7721 cells by RNA interference.In the first 4,5,6 days of proliferation experiment,the proliferation of interference group was significantly lower than the control group(separately F =17 098.922,18 135.107,32 641.075,all P ＜ 0.05 ); 48 h after transfection,the apoptosis rate of interference group was significantly higher than the control group (F =1136.452,P ＜0.05); In the infestation and migration experiments,the cells through the transwell chamber in the interference group were less than the control group( separately F =83.6,1126.857,all P ＜ 0.05 ) ; 24 h and 48 h after transfection,the vasculogenic mimicry in the interference group did not form which was significantly different from the control group. Conclusions Interfering the expression of nodal gene inhibits the malignant biological behaviour and the formation of vasculogenic mimicry in human hepatocellular carcinoma cells.

Objective To investigate the effect of all-trans retinoic acid (ATRA) on expression of gap junction genes 26, 32 and 43 in two human hepatocellular carcinoma cell lines. Method Human hepatocellular carcinoma cell lines SMMC-7721 and BEL-7404 were cultured and inoculated in culture plate. Cells were divided into 2 groups and cultured respectively with normal medium and medium contains ATRA at a concentration of 10-mol/L. After 24, 48 and 72 hours, cells were collected and total mRNA were extracted. RT-PCR was used to detect the transcription of gap junction genes 26, 32 and 43. Result SMMC-7721 and BEL-7404 cultured under condition of normal medium do not express CX26 mRNA and CX32 mRNA. With the effect of ATRA for 48 and 72 hours, SMMC-7721 cell expressed CX26 mRNA and CX32 mRNA. BEL-7404 expressed CX26 mRNA and CX32 mRNA while cultured with ATRA for 72 hours. CX43 mRNA expression of the two cell lines was not altered by ATRA. Conclusion ATRA affects the expression of CX26 and CX32 in HCC cell lines SMMC-7721 and BEL-7404 at the transcription level.

Objective To explore the prognostic factors of brain metastases from primary breast cancer treated with stereotactic radiotherapy (SRT).Methods Retrospectively analyze 37 brain metastatic patients from primary breast cancer.Among these patients nineteen were treated with stereotactic radiotherapy alone,eight patients were treated with whole brain radiotherapy (WBRT) plus SRT,the other ten patients were intracranial recurrence after WBRT and treated with SRT for salvage.Kaplan-Meier analyses were used to calculate survival time.Logrank and Cox proportional hazards regression analyses were performed for univariate and multivariate analyses.Results The median follow-up time were 11 months and 15 months for the whole group and the alive.The median overall survival time was 11 months (95％ CI =6-16.month) for the whole group.The median overall survival time was 13.5 months for the whole group.In univariate analysis,the triple negative breast cancer (x2 =5.95,P =0.004),lower Karnofsky performance score (KPS,x2 =13.85,P =0.000),the interval between the diagnosis of the primary tumor and brain metastases ≤ 30 months (x2 =6.62,P =0.010),high RPA grade (x2 =15.35,P =0.000) and intracranial recurrence after whole brain radiotherapy (x2 =4.43,P =0.035) were negative prognostic factors for brain metastasis of breast cancer treated with stereotactic radiotherapy.In multivariate analyses,the triple negative breast cancer (x2 =9.58,P =0.008),lower KPS (x2 =6.65,P =0.010),and intracranial recurrence after whole brain radiotherapy (x2 =3.95,P =0.047) were negative prognostic factor.Conclusion The triple negative breast cancer,lower KPS,and intracranial recurrence after WBRT were negative prognostic factor for brain metastasis from primary breast cancer treated with SRT.