Adult ; Cryptococcosis ; physiopathology ; Cryptococcus neoformans ; Duodenal Diseases ; microbiology ; physiopathology ; Humans ; Male

Objective To find out whether NOTCH receptors can serve as direct downstream targets of transforming growth factor β(TGF-β)/Smad4 signaling in endothelial cells.Methods Real-time PCR and Western blotting were performed to verify whether the expression of notch1 and notch4 was regulated by TGF-β pathway.Luciferase reporter assay was employed to investigate how the promoter of notch1 and notch4 was regulated by TGF-β.Then, ChIP assay was used confirm whether the promoter of notch1 and notch4 physically interacted with SMAD protein.Results TGF-β1 and bone morphogenetic protein 4 (BMP4) treatment increased the expression of both notch1 and notch4 at the transcriptional level.In addition, SMAD4 was physically associated with the SMAD binding sites on the notch4 promoter, which was largely enhanced under the treatment of TGF-β1 and BMP4.Importantly, TGF-β1 and BMP4 failed to transactivate notch4 in the absence of endogenous SMAD4 or the SMAD binding regions on the notch4 promoter.Conclusion The expression of NOTCH receptor can be directly up-regulated by SMAD4-mediated TGF-β/BMP signaling in cerebrovascular endothelial cells.

Objective To observe the expression of cAMP -response element binding protein (CREB) and N-methyl-D-aspartic acid receptor (NMDA ) after intracochlear electrical stimulation in the auditory cortex and inferior colliculus in infant rats with auditory deprivation .Methods Sixty six SD infant rats were randomly divided into 6 groups (11 rats each group):4 weeks ,and 6 weeks after injection of ototoxic drug ,the control group ,and 3 weeks and 5 weeks after injection of ototoxic drug with intra -cochlear electrical stimulation for one week .Gentami-cin sulphate (350 mg/kg body weight) and frusemide (200 mg/kg body weight) were injected subcutaneously in the skin folds on the lateral abdominal side and the dorsal neck area ,respectively .The expression of CREB and NMDAR1protein were detected by immunohistological staining .Results The results of immunohisto -chemistry revealed that protein expression of CREB and NMDAR1 in 4 week group of injection increased as compared to the control group ,while decreasing as compared to intracochlear electrical stimulation group ,significantly .However ,protein expression of CREB and NMDAR1 in 6 week group of injection decreased as compared to the control group and in-tracochlear electrical stimulation group ,significantly .Conclusion Auditory deprivation could result in the expres-sion of protein of CREB and NMDAR1 in auditory cortex and inferior colliculus increasing in an early stage and then de-creasing in infant rats .Intracochlear electrical stimulation could result in the expression of proteins of CREB and NMDAR 1 in auditory cortex and inferior colliculus increasing in infant rats .The dynamic variation of CREB and NMDAR1 expression in rat auditory cortex and inferior colliculus reflects synaptic plasticity in neurons of auditory pathway .

Objective To explore the clinical value of fetal syetem ultrasound union real-time three-dimensional ultrasound to diagnose the abnormalities of fetal palms and feet in medium-term pregnancy.Methods The results of fetal syetem ultrasound u-nion real-time three-dimensional ultrasound in 23 675 cases during dmedium-term pregnancy in our department from January 2009 to November 2013 were retrospectively analyzed,including 47 350 palms and feet.Results If using the fetal syetem ultrasound u-nion real-time three-dimensional ultrasound to examine fetal palms and feet more than three times,the display rate of palms and feet was 100.0%,while the first-time display rate of finger and toes was 81.2%,second-time display rate was 97.2% and the third-time and more display rate more thatn 99.8%.136 cases hand-foot deformity were diagnosed,including 37 cases of hand gesture abnor-malities,6 cases of finger abnormalities,93 cases of food abnormalities,and the main abnormality was strephexopodia.Of all the 136 cases,there were 2 cases also with Trisomy 18,4 cases with Trisomy 21.Conclusion Malformations of fetal palms and feet can be detected by fetal system ultrasound combined with real-time three-dimensional ultrasonography during the second trimester,which is important indicators of prenatal screening for chromosomal abnormalities.

To explore the molecular mechanism of human papillomavirus subtype 16(HPV-16)E7 oncogene-induced DNA re-replication in response to DNA damage. Flow cytometry was performed to examine the cell cycle changes in RPE1 E7 cells stably expressing HPV-16 E7 and its control cell RPE1 Vector after DNA damage.Immunoblotting assay was used to evaluate the early mitotic inhibitor 1(Emi1)expression in RPE1 E7 and RPE1 Vector cells with or without DNA damage.The changes of the proportion of polyploidy was detected by flow cytometry in DNA-damaged RPE1 E7 cells interfered by Emi1 small interfering RNA. Compared with the control cells,the proportion of polyploids in RPE1 E7 cells was significantly increased in response to DNA damage(=6.397,=0.0031).Emi1 protein expression was significantly increased in DNA damaged RPE1 E7 cells(=8.241,=0.0012).The polyploid ratio of RPE1 E7 cells was significantly reduced after Emi1 was interfered by two independent small interfering RNAs(=2.916,=0.0434;=3.452,=0.0260). In response to DNA damage,Emi1 promoted DNA re-replication caused by HPV-16 E7.
DNA Damage ; DNA Replication ; Human papillomavirus 16 ; Mitosis ; Oncogene Proteins, Viral

Background As one of the most common microvascular complication of diabetes in eyes,diabetic retinopathy (DR) is one of the most important cause of blindness.Nuclear factor-kappa B (NF-κB) is involved in the occurrence and development of the disease through the activation of a series of inflammatory cytokines.Objective The present study was to investigate the effects of peroxisome proliferator-activated receptor-gamma (PPAR-γ) excitomotor,rosiglitazone,on NF-κB expression and apoptosis of the retinal ganglion cells (RGCs) in the retina with diabetes mellitus. Methods Ninety SPF male Wistar rats were randomized into normal control group,diabetic control group and rosiglitazone group.Diabetes mellitus was induced by intraperitoneal injection of 50 mg/kg streptozotocin(STZ).Then 3 mg/kg rosiglitazone was intragastricly administered once per day in the rosiglitazonegroup,and the same volume of saline solution was used at the same way in the normal control group and diabetic control group from 3 days after modeling.The rats were sacrificed and the eye cups specimen was made at 4,8 and 12 weeks after usage of drugs.Retinal histopathological examination was performed by hematine-eosin staining,and expression of NF-κB p65 protein in retina and apoptotic index(AI) of RGCs were detected by immunohistochemistry and TUNEL assay,respectively in different time points mentioned above.The use of the animals complied with the Regulations for the Administration of Affairs Concerning Experimental Animals by State and Technology Commission.Results The blood glucose level was significantly elevated at various time points in the diabetic control group and rosiglitazone group compared with normal control group (P＜0.01 ),and that of the rosiglitazone group was significantly declined in comparison to the diabetic control group (q =0.81,0.82,1.23,P＞ 0.05 ).Normal retinal structure was seen in the normal control group,and edema retinal cell and disorder of retinal layers were exhibited in the diabetic control group.Retinal structure was almost normal in the rosiglitazone group.The NF-κB p65 was expressed weakly in the retina of normal control group,but the expression of NF-κB p65 was significantly elevated in the diabetic control group and rosiglitazone group compared with the normal control group(P＜0.01 ).However,the expression of NF-κB p65(A value)was significantly decreased in the rosiglitazone group compared with diabetic control group at 8 weeks and 12 weeks( q=17.77,15.30,P＜0.01 ).There were a few apoptotic cells in rat retina of the normal control group.Compared with the normal control group,the AI of the diabetic control group and rosiglitazone group was significantly reduced(P＜0.01 ).However,the AI of RGCs in the rosiglitazone group was significantly lower than that of diabetic control group in various time points (q =19.28,27.39,49.92,P＜0.01 ). Conclusions As one of the PPAR-γexcitomotors,rosiglitazone can inhibit apoptosis of RGCs through downregulating the expression of NF-κB in rat retina with diabetes mellitus,indicating a protective effect of rosiglitazone on retina in diabetic rat.

Objective To investigate the endothelial dysfunction in pre-hypertension and its influencing factors.Methods A total of 373 youth were divided as the subjects into hypertension group (HBP group),prehypertension group (PHT group) and normal blood pressure group (NBP group).Endothelial function was assessed based on carotid intima-media thickness (IMT),brachial artery flow-mediated dilation (FMD) and brachial-ankle pulse wave velocity (baPWV).Results IMT and baPWV in PHT group were higher than those in NBP group (P＜0.05),but did not reach the significant difference when compared with HBP group (P＞0.05).Compared with HBP,the levels of FMD in PHT group significantly increased (P＜ 0.05);however,no difference was observed in comparison with NBP group (P＞0.05).In the early stage of hypertension,diastolic BP (β=-0.120,P＜0.05) and body mass index (β=-0.115,P＜0.05) were negatively correlated with FMD;diastolic BP (β=0.146,P＜0.05),2-hour glucose (β=0.147,P＜0.05),high-density lipoprotein cholestrol (β=0.150,P＜0.05),and waist-hip ratio (β=0.126,P＜0.05) showed a positive correlation with IMT.baPWV was correlated with systolicBP (β=0.358,P＜0.01),waist circumference (β=0.254,P＜0.05),fasting glucose (β=0.155,P＜0.05),postprandial 2 h blood glucose (β =0.152,P ＜0.05),uric acid (β =0.206,P ＜ 0.05),and C-reactive protein (β=0.099,P＜0.05).Corclusion Our study shows that endothelial dysfunction may exist in the prehypertensive young,and several cardiovascular risks contribute to its development in the early stage of hypertension.

Background Our previous study demonstrated that epigallocateehin-gallate(EGCG),an active ingredient of green tea,has protective effect on optical nerve after optic nerve crush.Astrocyte was proved to play key role in the repair of nerve tissue,but the influence of EGCG on astrocyte is unclear.Glial flbrillary acidic protein (GFAP) is a special marker for astrocyte. Objective The aim of this study was to investigate the effect of EGCG on the expression of GFAP in optic nerve tissue after optic nerve crush. Methods Seventy-two clean Wistar rats were randomly divided into normal control group,sham+EGCG group,optic nerve crush+normal saline group(vehicle group),optic nerve crush+EGCG group.Optic nerve crush models were established by clamping optical nerve for 60 seconds by minitype optic nerve clipper with the force of 40 gram.Only ocular tissue was cut in the rats in sham group.Normal saline solution or EGCG(25 mg/kg)was intraperitoneally injected daily for 5 days consecutively and orally administered(2 mg/kg)daily afterwards.The expression of GFAP in optic nerve was detected by immunohistochemistry and quantified by Western blotting analysis on day 7,14 and 28 after modeling. Results lmmunochemistry showed that GFAP were weakly expressed in the rats of both normal group and sham+EGCG group with the sliSht brown staining in optic nerve tissue.The deeply brown staining for GFAP was seen in vehicle group,and the staining intensity weakened in optic nerve crush+EGCG group compared to vehicle group on days 7,14 and 28 after modeling.Western blotting analysis revealed that the expression level of GFAP in rat optic nerve tissue of vehicle group was significantly enhanced in comparison with normal control group(P＜0.01).On day 7 and 14 after optic nerve modeling,the expression levels of GFAP were evidently decreased in optic nerve crush+EGCG group in comparison with vehicle group(P＜0.05).However,on day 28 after modeling,no significant difference wag found in the expression levels of GFAP between vehicle group and optic nerve erush+EGCG group(P＞0.05). Conclusion EGCG down-regulates optic nerve crush-induced of GFAP in the optic nerve and therefore attenuates the activity of astrocytes,suggesting that EGCG might reduce the formation of glial scar.

Background Researches demonstrated that epigallocatechin-3-gallate(EGCG) can protect retinal ganglion cells(RGCs) against damage induced by retinal ischemia-reperfusion and optic nerve crush(ONC),but the effect of EGCG on lateral geniculate nucleus(LGN) was under study.Objective This study was designed to detect neuroprotective effect of EGCG on LGN in the rat model with ONC.Methods Forty-eight 7-week-old female clean Wistar rats were randomly divided into normal control group,sham operation+EGCG group,ONC+normal saline(NS) group and ONC+EGCG group.ONC models were created by clamping the optical nerve for 60 seconds with the clipper with the force of 40 grams in the right eyes of 24 rats.The EGCG solution(25mg/kg) was intraperitoneally injected from 2 days before operation daily for 5 consecutively days and orally administered(2mg/kg) after that,and NS was used in the same way for ONC+NS group.Four weeks after ONC,the brain tissue of the rats was obtained,and the neurons of dorsal LGN(dLGN) were counted by Nissl staining under the light microscopy.The expression of neurofilament triplet L(NF-L) was detected by immunohistochemistry and Western blot analysis.Meanwhile,the neuronal nitric oxide synthase(nNOS) positive cells were counted.Results Compared with normal control group,no significant differences were found in neuron number both between sham operation+EGCG group or ipsilateral LGN of operative eyes in ONC+normal saline group and ONC+EGCG group(P=0.906,0.561,0.794,0.646 respectively) in 4 weeks after ONC,but loss of neurons in contralateral LGN in both ONC+normal saline group and ONC+EGCG group were observed(P=0.000,0.015 respectively).However,compared with ONC+normal saline group,the density of neurons in ONC+EGCG group was higher(P=0.007).Moreover,a higher expression level of NF-L protein was seen in ONC+EGCG group compared with ONC+normal saline group at contralateral LGN of operative eyes(P=0.002).Concerning the number of nNOS positive cells in LGN,there was no significant difference among normal control group,sham operation+EGCG group and ONC+EGCG group(P＞0.05).The number of nNOS positive cells in the contralateral LGN of operative eyes of ONC+normal saline group was higher than that of ONC+EGCG group(P=0.000).Conclusion EGCG plays the protective effect on LGN after ONC in rats through mediating the expression of nNOS.

Background Vitreous hemorrhage in long-term produces toxic substances and influent the metabolism of eye tissue.Lens capsule is found more thin and transparent in the eyes with chronic vitreous hemorrhage.To research the effect of vitreous hemorrhage to lens is very important for the choose of the phaco operative timing.Objective The aim of this work was to investigate the ultrastructural change of lens epithelial cells(LECs) in the eyes with experimental vitreous hemorrhage.Methods The autologous blood of 0.1 ml was intravitreally injected in the left eyes of 8 general New Zealand white rabbits,and the equal amount of phosphate buffered saline(pH7.4) was used at the same way in the right eyes.Vitreous and fundus were examined with direct ophthalmoscope on 1,3,5,9,15,20,25,30 days to assess the inflammatory response after intravitreal injection.The specimens of lens anterior capsule were obtained in 30 days after injection and the ultrastructure and apoptosis of LECs were evaluated under the transmission electron microscope. Results No obvious ocular inflammatory response was seen throughout the experimental duration,and there was no vitreous hemorrhage in the right eyes after intravitreal injection.The vitreous hemorrhage agglutinated with clear boundary in the left eyes on 1 day after intravitreal injection,and the hemorrhage turned into dark-red color on the fifth day.On the fifteenth day after injection,the hemorrhage mass became to be grey color and the vitreous liquefaction occurred in the left eyes.The hemorrhage disappeared until 25 days.But in the one month after injection of self-blood,the vitreous showed the deeper red color.The early apoptosis appeared in the LECs of the left eyes in the thirty day,presenting the enlargement and broaden of intercellular space,the decrease of mitochondria number,vacuolar change,expanse of endoplasmic reticulum and disappearance of the nuclear membrane structure. Conclusions Vitreous hemorrhage leads to the ultrastructural pathological changes of lens.